RARE DISEASE
Infant botulism
Infant botulism
Infant botulism
Synonyms: Infant intestinal botulism, Infant intestinal toxemia botulism, Infant intestinal toxin-mediated botulism, Infantile botulism
Synonyms: Infant intestinal botulism, Infant intestinal toxemia botulism, Infant intestinal toxin-mediated botulism, Infantile botulism
Synonyms: Infant intestinal botulism, Infant intestinal toxemia botulism, Infant intestinal toxin-mediated botulism, Infantile botulism
Drug discovery
1
drug
With orphan designation
Overview
Infant botulism is a neuroparalytic disorder caused by intestinal colonization with Clostridium botulinum spores, which produce neurotoxins (A/B most common). It presents with constipation, hypotonia, cranial nerve deficits, and progressive flaccid paralysis, often requiring respiratory support. Early diagnosis (clinical suspicion ± stool toxin testing) and immediate treatment with botulism immune globulin (BabyBIG®) significantly reduce morbidity [1][6][10][16].
Therapies
BabyBIG® (antibody antitoxin): Reduces hospitalization from ~5.7 to ~2.3 weeks when administered early [3][10][16].
Supportive care: Ventilatory support (20–60% require intubation), enteral/parenteral nutrition, and monitoring for autonomic dysfunction [3][6][9].
Avoid aminoglycosides and clostridiocidal antibiotics, which may exacerbate paralysis [3][10].
Categories: rare infectious diseases, rare neurological diseases, rare ophthalmic disorders
Research Papers
717 drug discovery papers about Infant botulism, with 1 first-in-class and 4 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
717 drug discovery papers about Infant botulism, with 1 first-in-class and 4 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
small molecules
2026-06-02 | Botulinum neurotoxin: from molecular pathogenesis to emerging countermeasures.
Botulism is a severe neuroparalytic syndrome caused by botulinum neurotoxins (BoNTs)-among the most potent biological agents-with an estimated human lethal dose (LD50) of approximately 1 ng/kg. By cleaving soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins at cholinergic nerve terminals, BoNTs block acetylcholine release, producing potentially fatal flaccid paralysis. Despite advances in supportive care, current therapeutic options remain limited to antitoxin administration, which is effective only against circulating toxin and cannot neutralize intracellular BoNT. Consequently, patients often require prolonged mechanical ventilation and rehabilitation. To address these limitations, this review provides a comprehensive overview of botulism, spanning historical recognition, structural and mechanistic insights into BoNT activity, clinical manifestations, and current treatment strategies, while highlighting therapeutic gaps. Particular emphasis is placed on emerging preclinical interventions, including small-molecule inhibitors, antibody-based therapeutics, intracellular clearance strategies, and gene- and RNA-based modalities, reflecting rapid progress in structural biology and pharmacology. Collectively, these advances highlight both the promise and the remaining translational challenges of developing next-generation countermeasures, with implications for clinical management and biodefense preparedness.
2026-05-20 | Development of novel 2-[(8-hydroxyquinolin-7-yl)(phenyl)methylamino]benzoic acid derivatives for inhibiting the catalytic activity of botulinum neurotoxins type B and F: in silico, in vitro & in vivo evaluation.
Botulinum neurotoxins (BoNTs) are the most pernicious toxin and category 'A' bioterrorism agent, responsible for 'botulism', a rare but fatal disease. Toxins are divided into seven toxinotypes from A-G, where A, B, E, and F are accountable for human botulism. The available treatment strategies are inadequate and ineffective for post-neuronal intoxication. We report novel 2-[(8-hydroxyquinolin-7-yl)(phenyl)methylamino]benzoic acid derivatives targetingthe catalytic site of VAMP-acting serotypes (BoNT/B and F). Inhibitory potential of the synthesized compounds was studied using a framework of in silico and experimental approaches. Inhibitory activity and binding affinity were evaluated using substrate-based cleavage and SPR assays. Compound efficacy was tested in mice through pre-mixed, prophylactic, and therapeutic strategies. Molecular dynamics simulations analyzed binding interactions, structural fluctuations, and complex stability. Endopeptidase assay revealed that the selected compounds displayed ≥ 80% inhibition of BoNT/B and F catalytic activity, with IC50 values ranging from 17.58 to 34.05 µM. SPR analysis displayed binding affinity of these molecules ranging from 8.13E-06 to 7.69E-04 for both the proteins. In the mouse bioassay, the selected molecules displayed complete protection and extension in survival of up to 20-fold. MD simulation study supported the experimental finding, revealing key interactions with HExxH and other active site residues forming stable conformation throughout the simulation time. Among them, compounds A15 and A36 were predicted to be more effective inhibitors of BoNT/B and F serotypes, respectively. These findings could lay a promising way for the development of novel therapeutics by reducing disease severity, enhancing survivability, and recovery where no post-exposure therapy presently available.
2026-01-17 | Therapeutic potential of [(8-hydroxyquinolin-7-yl)(phenyl)methylamino] benzoic acid regioisomers against human-intoxicating botulinum neurotoxin serotypes: computational modeling to in vivo protection.
Botulinum neurotoxins are class I tier bioterrorism agent, accountable for causing rare but fatal illness ‘botulism’. Out of seven serotypes, A, B, E, and F are responsible for intoxicating humans. Despite knowing harmful effects on human health for centuries, there is no commercial antidote for post-neuronal intoxication is available. In the present study, we report efficacy of regioisomers of [(8-hydroxyquinolin-7-yl)(phenyl) methylamino]benzoic acid against zinc-dependent light chain activities of BoNT/A, /B, /E & /F by combining molecular modeling with in vitro and in vivo studies. Based on structure similarity search, multiple regioisomers of 8-hydroxyquinoline were mined and screened by performing molecular docking. The best-scored compounds were analyzed for inhibitory and binding potential against these serotypes via endopeptidase and surface plasmon resonance assays. The best two compounds (NSC1011 and NSC1012) with potential inhibition and binding kinetics across serotypes were evaluated for therapeutic potential in mouse model. NSC1011 and NSC1012 (regioisomers) docking data revealed their binding energies with active domains of BoNT/A, /B, /E, and /F light chains ranging between − 9.70 to − 4.27, and − 9.84 to − 7.23 kcal/mol, respectively. The endopeptidase assay displayed ˃ 90% inhibition of catalytic activities, with the IC50 values varying among serotypes from 20 to 40 µM concentrations. SPR interaction of both compounds with the targeted proteins was observed in the range of 3.83E-05 to 4.95E-04 M. These molecules have shown complete protection at one MLD (mouse lethal dose), whereas median extension of animal survival was recorded up to 24 h when exposed to 5X MLD. The in silico, in vitro, and in vivo data reveal that NSC1011 and NSC1012 exhibited good binding affinity, stability, inhibition with promising therapeutic potential against human botulism-causing toxinotypes.
2025-11-12 | Two Cases of Severe Food-Borne Botulism: Could High-Dose Methylcobalamin Accelerate Recovery?
Botulinum neurotoxins (BoNTs) are zinc-dependent endopeptidases that cleave presynaptic SNARE proteins and arrest acetylcholine release, producing flaccid paralysis with protracted recovery. We report two cases of a mother and son who developed severe food-borne botulism after eating Fesikh (traditional salted fish) with rapidly progressive descending paralysis requiring ventilation, prominent dysautonomia (hypotension responsive to fluids, bradycardia, dry mucosae, constipation with abdominal distension, as well as urinary retention), and intact consciousness. Both received trivalent antitoxin on day 2; the son was extubated on day 10, and the mother on day 16. Persistent bladder atony, fatigue, weakness, and poor concentration improved with structured physiotherapy accompanied by intramuscular methylcobalamin, which was initiated on day 1 of admission and subsequently transitioned to an eight-week course of oral methylcobalamin (500 µg daily) during the post-extubation rehabilitation phase. We discuss mechanistic plausibility and hypothesize that higher-dose methylcobalamin might accelerate neuromuscular recovery by stabilizing mitochondria, attenuating caspase-mediated apoptosis, and promoting axonal regeneration and sprouting.
2025-10-14 | Phosphorylation of SNAP-25 at Ser187 is enhanced following its cleavage by Botulinum Neurotoxin Serotype A, promoting the dominant-negative effect of the resulting fragment.
Botulinum Neurotoxin Serotype A (BoNT/A), responsible for most human botulism cases, inhibits neurotransmitter release by cleaving the target protein SNAP-25. Previous literature demonstrated that BoNT/A mediated cleavage of a small subset of the SNAP-25 pool, resulting in SNAP-25 (1-197) fragments, is sufficient to block exocytosis. SNAP-25 (1-197) potentially competes against intact SNAP-25 for SNARE complexes and blocks neurotransmission through a dominant-negative mechanism. However, how a tiny fraction of cleaved SNAP-25 efficiently outcompetes a large pool of intact SNAP-25 remains unknown. Here, we examined the importance of SNAP-25 phosphorylation at Ser187 residue, located in the C-terminus SNARE domain, in the context of BoNT action. Our results demonstrated that Ser187-phosphorylated SNAP-25 can be efficiently cleaved in cells. Importantly, BoNT/A-cleaved SNAP-25 fragments in neuronal and non-neuronal cells are heavily phosphorylated at Ser187 and localized on the cell membrane. SNAP-25 (1-197) binds to syntaxin-1A, and the interaction is enhanced by Ser187 phosphorylation. We also found that SNAP-25 (1-197) survives longer than the BoNT/A enzymatic component itself in cells. Molecular modeling suggested that SNAP-25 (1-197), phosphorylated or not, forms stable SNARE complexes; however, Ser187 phosphorylation induces local changes in surface electrostatic potential and dynamics of the complex. This study characterizes the molecular mechanism underlying the dominant-negative effect of SNAP-25 (1-197) on neurotransmission. This research could have implications for the future development of BoNT/A inhibitors and the generation of new BoNT/A clinical formulations by regulating the abundance of Ser187 phosphorylation in cleaved SNAP-25 fragments.
proteins
2026-03-06 | Investigating the dual nature of Clostridium botulinum: pathogenic mechanisms and therapeutic potentials.
Clostridium botulinum (C. botulinum) is known for producing one of the strongest neurotoxins, namely botulinum neurotoxin (BoNT). This toxin is responsible for causing botulism, which is considered a severe neurological disease. This comprehensive narrative review highlights the dual nature of C. botulinum as both a serious health threat and a groundbreaking therapeutic agent. Methods: A comprehensive literature search was conducted in Google Scholar, PubMed/Medline, and Scopus (1971-2025, 2020-2025 literary focus) using keywords related to botulism pathogenesis, clinical features, and therapeutic approaches. Key findings indicate that, despite advancements in botulism management and reduced mortality due to heptavalent botulinum antitoxin (HBAT) and botulism immune globulin intravenous (BIG-IV) antitoxins, diagnostic challenges and bacterial spore resistance, particularly in group I, remain persistent. On the other hand, the discovery of serotype BoNT/X with lower toxicity in vertebrates and unique capabilities has opened new horizons in the design of safer treatments. C. botulinum represents a remarkable paradox in biology, an organism capable of producing one of the deadliest toxins known, yet simultaneously a cornerstone in modern therapeutic innovation. In fact, this toxin's dual nature is based on its selective targeting of BoNTs. In this regard, the systemic entry and distribution of this poison in the body - or, in other words, intoxication with it - poses a life-threatening aspect. On the other hand, the use of this bacterium's toxin for therapeutic and medicinal purposes is progressing due to its topical and controlled administration and entry into the body make it a targeted and effective therapeutic tool in the field of medicine. A better and more precise understanding of the mechanisms of action and, if necessary, future structural and engineered modifications in the toxin could lead to the emergence of a new generation of BoNTs that hold promise for the development of esthetic and therapeutic applications in medicine.
2026-01-27 | Advances in Clostridial Neurotoxins: Passage of the Intestinal Barrier and Targeting of Specific Neuronal Cells.
Clostridial neurotoxins, botulinum neurotoxins (BoNTs), and tetanus neurotoxin (TeNT) are potent toxins responsible for severe diseases, botulism and tetanus, respectively. BoNTs associate with non-toxic proteins (non-toxic non-hemagglutinin, hemagglutinins, and OrfXs), which protect BoNTs against acidic pH and protease degradation and facilitate BoNT passage through the intestinal barrier. TeNT enters motor neurons and undergoes a retrograde axonal transport until the target inhibitory interneurons in the central nervous system. BoNTs and TeNT recognize specific cell surface receptors which consist of complex sets of protein(s)-glycan-gangliosides and determine specific cell entry pathways. Recent data on structural and functional investigations of BoNT and TeNT receptors bring a better understanding of toxin trafficking in the host and entry into target neuronal cells, which is useful for the development of updated strategies of prevention and treatment of the corresponding diseases. Since clostridial neurotoxins, notably BoNTs, are important therapeutic tools, detailed knowledge of their activity opens the way of the development of engineered molecules for specific clinical applications.
2025-06-05 | Botulinum neurotoxin A mutants with enhanced ganglioside binding show improved potency and altered ganglioside selectivity.
Botulinum neurotoxins are the causative agents of botulism, a lethal paralytic disease, but are also one of the most commonly used therapeutics for the treatment of numerous neuromuscular conditions. These toxins recognise motor nerve terminals with high specificity and affinity by using a dual binding mechanism involving gangliosides and protein receptors. The initial recognition of gangliosides is crucial for the toxins' potency. In this study, we employed a synaptosome-binding screening strategy to identify BoNT/A mutants with enhanced ganglioside-binding which translated into improved potency. X-ray crystallography and receptor-binding assays were used to elucidate the molecular mechanisms underlying the increased affinity or altered ganglioside selectivity of these mutants. Our findings provide a basis for the development of BoNT/A variants with enhanced therapeutic potential.
2025-04-23 | Recombinant Detoxified Holotoxin as a Potent Candidate Vaccine Against Botulism.
Vaccination may represent a suitable strategy for preventing botulism. The recombinant expression of toxin functional domains can induce effective immune responses against botulism. This study aims to develop a safe and effective recombinant detoxified full-length BoNTA vaccine. In this study, we engineered and mutated the toxin activity-related sites on the basis of the full-length BoNTA protein and constructed three detoxified full-length toxin mutants. They were recombinant expressed and purified in Escherichia coli, and the BoNTA/M4 mutant was determined to have the highest safety, with a murine lethal dose of 50% (MLD50). The M4 protein was used as the antigen for three immunizations, and the serum titers, neutralizing activity, and BoNTA protective effects of immunized mice were evaluated. The results show that, in comparison to the receptor-binding domain BoNTA/Hc protein, the full-length detoxified mutant M4 protein exhibited superior immunogenicity and could induce higher levels of specific antibodies, and the resulting immune serum could effectively protect mice against higher doses of BoNTA challenge. This study laid the foundation for research on a novel recombinant detoxified full-length botulinum toxin vaccine.
2025-01-24 | Development of a Recombinant Fusion Vaccine Candidate Against Lethal Clostridium botulinum Neurotoxin Types A and B.
Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are potent protein toxins that can cause botulism, which leads to death or neuroparalysis in humans by targeting the nervous system. BoNTs comprise three functional domains: a light-chain enzymatic domain (LC), a heavy-chain translocation domain (HCN), and a heavy-chain receptor-binding domain (HCC). The HCC domain is critical for binding to neuronal cell membrane receptors and facilitating BoNT internalization via endocytosis. Accordingly, it may serve as a vaccine candidate, inducing anti-BoNT-neutralizing antibodies in animals. Here, we aimed to develop a vaccine capable of simultaneously defending against both BoNT/A and B. We combined the HCC domains of botulinum neurotoxin type A (BoNT/A) and botulinum neurotoxin type B (BoNT/B) in Escherichia coli to produce a recombinant protein (rHCCB-L-HCCArHCcB) that offers dual protection against both toxins by inhibiting their receptor binding. To evaluate the efficacy of the vaccine, mice were immunized intramuscularly with rHCCB-L-HCCA plus alum thrice at 2-week intervals, followed by the assessment of immunogenicity and protective efficacy. The antibody titer in mice immunized with rHCCB-L-HCCA was significantly higher than that in mice immunized with alum alone, protecting them from the lethal challenges of BoNT/A (105 50% lethal dose, LD50) and B (103 LD50). These findings suggest that rHCCB-L-HCCA may simultaneously be an effective vaccine candidate against BoNT/A and B.
antibodies
2026-06-17 | When BAT Precedes BIG-IV in Infant Botulism: The Need for Structured Antitoxin Decision-Pathway Reporting in Case Reports.
In infant botulism, use of equine-derived botulinum antitoxin before BIG-IV may be clinically justified when the infant-specific product is delayed. Future case reports should document the treatment decision pathway, antitoxin timing, risk-benefit justification, guardian counseling, adverse-event monitoring, and relapse-prevention rationale to make such emergency decisions reproducible.
2025-12-12 | Infant Botulism: A Case Study in Integrated Clinical and Public Health Response.
Early recognition of constipation, feeding difficulty, and descending weakness in infants is vital for prompt diagnosis of infant botulism. Bedside nerve studies support early treatment. Rapid public health notification and coordinated access to specific therapies, including timely use of BAT and BIG-IV, are essential to optimize outcomes and safety.
2025-12-12 | Whole-genome sequencing-based typing methods for Clostridium butyricum strains from clinical, animal, plant, and environmental sources.
Clostridium butyricum exhibits a dual role, acting not only as a probiotic but also as an opportunistic pathogen associated with neonatal necrotizing enterocolitis (NEC) and infant botulism. We aimed to establish high-resolution genotyping frameworks to improve molecular surveillance and outbreak investigations. We analyzed 297 C. butyricum genomes, including 200 isolates from preterm neonates across 13 French neonatal intensive care units over a 20-year period and 97 publicly available genomes. A core-genome multilocus sequence typing (cgMLST) scheme was developed using chewBBACA, defining 2,621 loci, and applied to genomes with ≥95% locus presence. Core-genome single-nucleotide polymorphism (cgSNP) analysis was performed for complementary resolution. Phylogenetic cgMLST classified isolates into nine major clades. Some clinical strains displayed clonal relationships, whereas others were geographically and temporally unrelated. All botulinum neurotoxin type E-producing strains were grouped within a single clade. NEC-associated isolates showed geographic and temporal clustering, but no clade was uniquely linked to NEC. cgSNP analysis identified 11 clusters with overall discriminatory power similar to cgMLST while providing finer resolution for NEC-related strains. We propose robust cgMLST and cgSNP schemes for C. butyricum, enabling high-resolution genotyping and supporting epidemiological surveillance and outbreak investigation of this emerging opportunistic pathogen in neonatal settings. Clostridium butyricum has been identified in fecal samples from both asymptomatic neonates and cases of necrotizing enterocolitis (NEC). Using a large collection of strains from different origins and spatiotemporal contexts, we developed and established a cgMLST scheme for the molecular typing of C. butyricum. Our results show that most C. butyricum strains cluster independently of origin and spatiotemporal context factors. However, specific cgMLST clades of C. butyricum were found for plant and botulinum neurotoxin type E strains. Clonal strains were also identified. No specific cgMLST clade was found to be genetically associated with NEC. cgSNP showed higher discriminatory power compared to cgMLST. Importantly, cgSNP provided better discriminatory power for strain relatedness with respect to strains isolated from NEC patients.
2025-11-26 | Regional Risk Discrepancy Indicates a Slow Outbreak of Infant Botulism, Victoria, Australia.
Infant botulism accounts for many new botulism cases each year. Geographic case clustering has been previously described in California. This study aimed to describe whether case clustering was present in Victoria, Australia. We conducted a retrospective case series of infant botulism presenting to Victorian hospitals between 1978 and 2024. Moran's I test for spatial autocorrelation was used to determine if spatiotemporal clustering was present. Twelve cases of infant botulism presented to Victorian hospitals in the study period. Using Moran's I statistic in analyzing the number of cases in each postcode of interest, a case clustering was demonstrated in the northwest of Victoria ( P < 0.01). We describe a geographic case clustering within the relatively sparsely populated northwest in Victoria, Australia.
2025-09-26 | Features of the First Case of Foodborne Botulism Caused by Dual-Toxin Clostridium parabotulinum Subtype A1(B5) in Spain.
The neurotoxin BoNT/B2 is the predominant Clostridium parabotulinum subtype in foodborne and infant botulism cases in Spain. This study characterizes a novel case of foodborne botulism in Spain caused by a dual-toxin A1(B5) strain. A 64-year-old male presented with acute, progressive flaccid paralysis including diplopia, dysphagia, and respiratory failure. Although botulism was not initially suspected, the patient recovered with supportive care and without antitoxin administration. Genomic characterization confirmed the presence of both bont/A1 and silent bont/B5 genes. The bont/A1 gene was associated with an orfX+ neurotoxin gene cluster, while the silent bont/B5 gene was in an ha+ cluster. Phylogenetic analysis of both bont/A1 and bont/B5 sequences showed 100% amino acid identity, respectively, to previously reported A1(B5) strains (e.g., CDC_69094, FE9504ACG). Multi-locus sequence typing (MLST) assigned the ST10, a genotype previously undetected in Spanish botulism cases, yet found in other European countries. This case highlights the importance of considering botulism in differential diagnosis due to its varied presentation and the significance of timely laboratory confirmation for effective management. The identification of this dual-toxin BoNT/A1(B5) orfX+/ha+ ST10 strain expands our understanding of C. botulinum epidemiology and genetic diversity in Spain.
vaccines
2026-01-27 | Proteolytic Activities and Immunological Effects of Light Chains of Botulinum Neurotoxin A1, A2 and A3 Subtypes.
Botulinum neurotoxin serotype A (BoNT/A) is the most potent known neurotoxin. While its light chain (LC) catalytic domain is a prime target for next-generation vaccines and therapeutics, the functional differences among BoNT/A subtype LCs (A1, A2, A3) remain to be definitively characterized, despite notable sequence variation. This work aimed to systematically compare the proteolytic activity and immunoprotective efficacy of recombinant BoNT/A1-LC, A2-LC, and A3-LC. Recombinant A1-LC-His, A2-LC-His, A3-LC-His, and A3-LC-Twin-Strep proteins were expressed in Escherichia coli (E. coli) and purified with affinity chromatography. Their proteolytic activity was assessed via in vitro SNAP-25 cleavage assays. The protective potency of these antigens was evaluated in a mouse model. In vitro cleavage assays revealed a substrate cleavage efficiency order of A2-LC > A1-LC > A3-LC. In vivo, both A1-LC and A2-LC immunization conferred robust, broad protection against high-dose challenges with all three toxin subtypes. In stark contrast, A3-LC provided only minimal protection against its homologous toxin and none against heterologous subtypes. Crucially, the functional deficit of A3-LC was confirmed to be an intrinsic property, as the A3-LC-TS variant, designed to exclude tag-specific interference, exhibited comparable low efficacy. According to structural research, A3-LC's compromised function may be caused by a four-amino-acid loss. The inferior performance of A3-LC is inherent to its primary structure. This work identified A1-LC or A2-LC as the potential proteolytic activity molecule and vaccine antigen by demonstrating functional differences among BoNT/A subtype LCs. These findings provide crucial insights for developing subtype-specific countermeasures against botulism.
2025-12-16 | Self-Applicable Thermostable Heterogeneous Microneedle Patch for Recombinant Protein Vaccination against Botulinum Neurotoxin Serotype A.
Botulinum neurotoxin serotype A (BoNT/A), a Class A bioterrorism agent, poses significant poisoning risks, with diagnosis challenges and no specific antidotes, making vaccination the most effective prevention. However, traditional needle-based vaccines are painful, thermally unstable, require trained professionals for administration, and carry infection risks, leading to low patient compliance. To overcome these limitations, we introduce a self-applicable, thermostable, heterogeneous microneedle (MN) patch for delivering a recombinant protein vaccine based on the C-terminal heavy chain of BoNT/A to prevent botulism. By incorporating materials with varying hydrophilicity, the MN patch enhances drug-layer contact with skin, improving vaccine delivery efficiency. Fabricated via heat- and pressure-assisted micromolding, the polycaprolactone MNs demonstrate sufficient mechanical strength for effective skin penetration. In vivo studies in a mouse model show that MN patches loaded with the recombinant C-terminal heavy chain of BoNT/A elicit strong IgG antibody responses and provide protective efficacy comparable to those of intramuscular injections, with all immunized mice surviving lethal BoNT/A challenges. The IgG responses of MN patches did not decrease after storage for 6 mol (25 °C, 60% relative humidity), indicating good stability. These findings suggest that thermostable MN patches offer a noninvasive, effective, patient-friendly alternative to traditional vaccines, with potential applications in bioterrorism prevention.
2025-04-18 | Detoxification techniques for bacterial toxins: A pathway to effective toxoid vaccines.
Bacterial toxins play a critical role in the virulence of many pathogens, leading to serious diseases such as tetanus, diphtheria, botulism, and entrotoxemia. As key virulence factors, these toxins cause significant tissue damage and disease manifestations in infected hosts. Vaccination against these toxins through toxoid vaccines, composed of inactivated forms of the toxins, represents a vital strategy for preventing toxin-mediated diseases. However, creating effective toxoid vaccines necessitates meticulous detoxification processes that ensure the loss of toxicity while retaining the immunogenic properties inherent in the native toxins. This review offers a comprehensive evaluation of the diverse methodologies employed for detoxifying bacterial toxins, highlighting their advantages, limitations, and implications for vaccine development. By detailing comparisons of efficacy, stability, residual toxicity, and clinical applicability, we demonstrate that while traditional methods utilizing chemical reagents (such as formaldehyde) remain widely used, emerging technologies like genetic inactivation and protein engineering present significant advantages. These innovations promise to advance the development of durable and irreversible toxoid vaccines that protect public health and contribute to future vaccine formulation improvements. Ultimately, this knowledge synthesis aims to guide future research efforts and facilitate the design of safer and more effective toxoid vaccines to combat the public health threats posed by toxin-producing bacteria.
2024-10-02 | New Engineered-Chimeric Botulinum Neurotoxin Mutant Acts as an Effective Bivalent Vaccine Against Botulinum Neurotoxin Serotype A and E.
Botulinum neurotoxins (BoNTs), including serotypes A and E, are potent biotoxins known to cause human poisoning. In addition to the critical protective antigen found in the full BoNT molecule, the receptor binding domain (Hc domain), BoNTs also harbour another essential protective antigen-the light chain-translocation domain (L-HN domain). Leveraging these pivotal protective antigens, we genetically engineered a series of inactivated chimeric molecules incorporating L-HN and Hc domains of BoNT/A and E. The structure of these chimeric molecules, mirror BoNT/A and E, but are devoid of enzyme activity. Experimental findings demonstrated that a lead candidate mEL-HN-mAHc harnessing the inactivated protease LCHN/E with the mutated gangliosides binding site Hc/A (mE-mA) elicited robust immune protection against BoNT/A and E simultaneously in a mouse model, requiring low immune dosages and minimal immunisations. Moreover, mE-mA exhibited high protective efficacy against BoNT/A and E in guinea pigs and New Zealand white rabbits, resulting in elevated neutralising antibody titres. Furthermore, mE-mA proved to be a more stable and safer vaccine compared to formaldehyde-inactivated toxoid. Our data underscore the genetically engineered mE-mA as a highly effective bivalent vaccine against BoNT/A and E, paving the way for the development of polyvalent vaccines against biotoxins.
2024-09-24 | Novel platform for engineering stable and effective vaccines against botulinum neurotoxins A, B and E.
Botulinum neurotoxin (BoNT), produced by Clostridium botulinum, is the most toxic protein known, capable of causing severe paralysis and posing a significant bioterrorism threat due to its extreme lethality even in minute quantities. Despite this, there are currently no FDA-approved vaccines for widespread public use. To address this urgent need, we have developed an innovative vaccine platform by fusing the neuronal binding domain of BoNT/E (Hc/E) with core-streptavidin (CS), resulting in a stable CS-Hc/E vaccine. Mice vaccinated with CS-Hc/E exhibited superior antibody titers compared to those receiving Hc/E alone. To develop a trivalent vaccine against BoNT/A, BoNT/B, and BoNT/E- key contributors to the vast majority of human botulism-we conjugated CS-Hc/E with a biotinylated atoxic chimeric protein incorporating neutralizing epitopes from BoNT/A and BoNT/B. This chimeric protein includes the binding domain of BoNT/A, along with the protease-inactive light chain and translocation domains of BoNT/B. The interaction between CS and biotin formed a stable tetrameric antigen, EBA. Vaccination with EBA in mice elicited robust antibody responses and provided complete protection against lethal doses of BoNT/A, BoNT/B, and BoNT/E. Our findings highlight EBA's potential as a stable and effective broad-spectrum vaccine against BoNT. Moreover, our technology offers a versatile platform for developing multivalent, stable vaccines targeting various biological threats by substituting the BoNT domain(s) with neutralizing epitopes from other life-threatening pathogens, thereby enhancing public health preparedness and biodefense strategies.
other
2026-06-24 | Acquisition of Stickland-metabolizing bacteria during infancy prevents Clostridium botulinum infection
Summary The mechanisms by which maturation of the infant gut microbiota promotes resistance to pathogen colonization remain poorly understood. Infant botulism, a potentially fatal disease caused by intestinal colonization by Clostridium botulinum , provides a striking example of age-dependent susceptibility to infection 1 . Although the gut microbiota has long been implicated in protection against C. botulinum 2–6 , the responsible bacterial species and underlying mechanisms have yet to be elucidated. Here, we show that acquisition of Stickland-metabolizing Clostridia during infant gut microbiota maturation confers resistance to C. botulinum colonization through competition for shared amino-acid-dependent nutritional niches. In human fecal microbiota-transplanted mice, longitudinally collected infant microbiotas exhibited a clear transition from susceptibility to resistance. Intestinal metabolomic analysis identified 5-aminovalerate as a hallmark metabolite of the resistant microbiota, implicating Stickland metabolism, an amino acid metabolic pathway also utilized by C. botulinum . Guided by this finding, metagenomic analysis revealed enrichment of Stickland-metabolizing Clostridia in resistant microbiotas, including Clostridioides difficile , a bacterium frequently carried by healthy infants. Intestinal metabolic signatures of C. botulinum and infant-derived Stickland-metabolizing Clostridia suggested competition for shared amino-acid-dependent nutritional niches. Consistent with this model, C. difficile suppressed C. botulinum expansion through nutrient competition. Together, these findings identify nutritional niche competition as a mechanism by which microbiota maturation promotes resistance to C. botulinum colonization. This work demonstrates how acquisition of specific microbial metabolic functions during early life can shift the gut microbiota from a susceptible to a resistant state.
2026-02-12 | Endemic koloa maoli (Hawaiian Duck, Anas wyvilliana ) shows preferential social associations, but not based on plumage or genetic relatedness.
Island endemic birds are under greater threat than their mainland counterparts. Sedentary living and historically reduced predation may affect island bird sociality and inform their conservation and management. However, detailed studies on their sociality are uncommon. The federally- endangered koloa maoli ( Anas wyvilliana, Hawaiian duck, or koloa) is primarily threatened by hybridization with feral mallards and avian botulism outbreaks. We used capture-mark-recapture and genetic data on koloa on the island of Kaua'i, a stronghold of remaining koloa, to construct social networks and examine their associations (inferred from co-occurrence in traps) and spatial genetic structure. Information on associations might shed light on preferences for or against mallards and hybrids, and inform planned translocation efforts. Microgeographic spatial genetic structuring where relatedness among individuals scales with geographic distance is a potential liability for maintaining koloa genetic diversity, and would particularly be detrimental during highly localized outbreaks of botulism that could result in the removal of entire lineages. While we found persistent social associations among adult koloa, they were not apparently influenced by plumage traits or body mass, suggesting a lack of social preference for mallard-like individuals. Importantly, we found no spatial patterns of relatedness within the largest refuge. Therefore, botulism outbreaks remain a demographic threat but are unlikely to remove correlated genetic diversity. There were no sex differences in spatial genetic structure and both sexes moved within a refuge. The lack of spatial genetic structure and the presence of many unrelated conspecifics may enable koloa to limit inbreeding and retain genetic diversity without sex-biased dispersal. In the context of future translocations, our results suggest that translocating koloa captured in the same trap together will reduce disruption of preferred associations while also retaining genetic diversity among translocated individuals. The koloa maoli (Hawaiian duck, or koloa) is a federally-endangered, island endemic bird. Like other Hawaiian waterbirds, koloa are threatened by introduced predators and habitat loss, but also specifically by hybridization with feral mallards and localized avian botulism outbreaks. Currently, the island of Kaua'i has the largest population of non-admixed koloa. We used capture-mark-recapture and genetic data of koloa at two wetland sites on Kaua'i to examine their sociality and spatial genetic structure.Koloa formed preferential social associations, but they were not based on plumage traits, body mass or genetic relatedness.There was no spatial genetic structure for males and females within a wetland site. Our results suggested that 1) koloa have no preference for mallard-like plumage that might increase hybrid pairing, 2) localized (within-refuge) botulism outbreaks are unlikely to remove close relatives and unique genetic lineages, and 3) translocation of groups could maintain social associations without limiting genetic diversity.
2024-01-02 | Development and characterization of a novel neutralizing scFv vectored immunoprophylaxis against botulinum toxin type A.
Botulinum toxin is a protein toxin secreted by Clostridium botulinum that is strongly neurotoxic. Due to its characteristics of being super toxic, quick acting, and difficult to prevent, the currently reported antiviral studies focusing on monoclonal antibodies have limited effectiveness. Therefore, for the sake of effectively prevention and treatment of botulism and to maintain country biosecurity as well as the health of the population, in this study, we intend to establish a single chain antibody (scFv) targeting the carboxyl terminal binding functional domain of the botulinum neurotoxin heavy chain (BONT/AHc) of botulinum neurotoxin type A, and explore the value of a new passive immune method in antiviral research which based on adeno-associated virus (AAV) mediated vector immunoprophylaxis (VIP) strategy. The scFv small-molecular single-chain antibody sequenced, designed, constructed, expressed and purified by hybridoma has high neutralising activity and affinity level, which can lay a good foundation for the modification and development of antibody engineering drugs. In vivo experiments, AAV-mediated scFv engineering drug has good anti-BONT/A toxin neutralisation ability, has advantages of simple operation, stable expression and good efficacy, and may be one of the effective treatment strategies for long-term prevention and protection of BONT/A botulinum neurotoxin.
2022-10-30 | Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A
ABSTRACT Single-domain antibodies (sdAbs, VHHs, or nanobodies) are a promising tool for the treatment of both infectious and somatic diseases. Their small size greatly simplifies any genetic engineering manipulations. Such antibodies have the ability to bind hard-to-reach antigenic epitopes through long parts of the variable chains, the third complementarity-determining regions (CDR3s). VHH fusion with the canonical immunoglobulin Fc fragment allows the Fc-fusion single-domain antibodies (VHH-Fc) to significantly increase their neutralizing activity and serum half-life. Previously we have developed and characterized VHH-Fc specific to botulinum neurotoxin A (BoNT/A), that showed a 1000-fold higher protective activity than monomeric form when challenged with five times the lethal dose (5 LD 50 ) of BoNT/A. During the COVID-19 pandemic, mRNA vaccines based on lipid nanoparticles (LNP) as a delivery system have become an important translational technology that has significantly accelerated the clinical introduction of mRNA platforms. We have developed an mRNA platform that provides long-term expression after both intramuscular and intravenous application. The platform has been extensively characterized using firefly luciferase (Fluc) as a reporter. An intramuscular administration of LNP-mRNA encoding VHH-Fc antibody made it possible to achieve its rapid expression in mice and resulted in 100% protection when challenged with up to 100 LD 50 of BoNT/A. The presented approach for the delivery of sdAbs using mRNA technology greatly simplifies drug development for antibody therapy and can be used for emergency prophylaxis.
2022-06-30 | Genomic and Phenotypic Characterization of Clostridium botulinum Isolates from an Infant Botulism Case Suggests Adaptation Signatures to the Gut.
In early life, the immature human gut microbiota is prone to colonization by pathogens that are usually outcompeted by mature microbiota in the adult gut. Colonization and neurotoxin production by a vegetative Clostridium botulinum culture in the gut of an infant can lead to flaccid paralysis, resulting in a clinical outcome known as infant botulism, a potentially life-threatening condition. Beside host factors, little is known of the ecology, colonization, and adaptation of C. botulinum to the gut environment. In our previous report, an infant with intestinal botulism was shown to be colonized by neurotoxigenic C. botulinum culture for 7 months. In an effort to gain ecological and evolutionary insights into this unusually long gut colonization by C. botulinum, we analyzed and compared the genomes of C. botulinum isolates recovered from the infant feces during the course of intoxication and isolates from the infant household dust. A number of observed mutations and genomic alterations pinpointed at phenotypic traits that may have promoted colonization and adaptation to the gut environment and to the host. These traits include motility, quorum-sensing, sporulation, and carbohydrate metabolism. We provide novel perspectives and suggest a tentative model of the pathogenesis of C. botulinum in infant botulism. IMPORTANCE While the clinical aspects of infant botulism and the mode of action of BoNT have been thoroughly investigated, little is known on the pathogenesis and adaptive mechanisms of C. botulinum in the gut. Here, we provide for the first time a comprehensive view on the genomic dynamics and plasticity of C. botulinum over time in a case of infant botulism. The genomic and phenotypic analysis of C. botulinum isolates collected during the disease course offers an unprecedented view of C. botulinum ecology, evolution, and pathogenesis and may be instrumental in developing novel strategies for prevention and treatment of toxicoinfectious botulism.
small molecules
2026-06-02 | Botulinum neurotoxin: from molecular pathogenesis to emerging countermeasures.
Botulism is a severe neuroparalytic syndrome caused by botulinum neurotoxins (BoNTs)-among the most potent biological agents-with an estimated human lethal dose (LD50) of approximately 1 ng/kg. By cleaving soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins at cholinergic nerve terminals, BoNTs block acetylcholine release, producing potentially fatal flaccid paralysis. Despite advances in supportive care, current therapeutic options remain limited to antitoxin administration, which is effective only against circulating toxin and cannot neutralize intracellular BoNT. Consequently, patients often require prolonged mechanical ventilation and rehabilitation. To address these limitations, this review provides a comprehensive overview of botulism, spanning historical recognition, structural and mechanistic insights into BoNT activity, clinical manifestations, and current treatment strategies, while highlighting therapeutic gaps. Particular emphasis is placed on emerging preclinical interventions, including small-molecule inhibitors, antibody-based therapeutics, intracellular clearance strategies, and gene- and RNA-based modalities, reflecting rapid progress in structural biology and pharmacology. Collectively, these advances highlight both the promise and the remaining translational challenges of developing next-generation countermeasures, with implications for clinical management and biodefense preparedness.
2026-05-20 | Development of novel 2-[(8-hydroxyquinolin-7-yl)(phenyl)methylamino]benzoic acid derivatives for inhibiting the catalytic activity of botulinum neurotoxins type B and F: in silico, in vitro & in vivo evaluation.
Botulinum neurotoxins (BoNTs) are the most pernicious toxin and category 'A' bioterrorism agent, responsible for 'botulism', a rare but fatal disease. Toxins are divided into seven toxinotypes from A-G, where A, B, E, and F are accountable for human botulism. The available treatment strategies are inadequate and ineffective for post-neuronal intoxication. We report novel 2-[(8-hydroxyquinolin-7-yl)(phenyl)methylamino]benzoic acid derivatives targetingthe catalytic site of VAMP-acting serotypes (BoNT/B and F). Inhibitory potential of the synthesized compounds was studied using a framework of in silico and experimental approaches. Inhibitory activity and binding affinity were evaluated using substrate-based cleavage and SPR assays. Compound efficacy was tested in mice through pre-mixed, prophylactic, and therapeutic strategies. Molecular dynamics simulations analyzed binding interactions, structural fluctuations, and complex stability. Endopeptidase assay revealed that the selected compounds displayed ≥ 80% inhibition of BoNT/B and F catalytic activity, with IC50 values ranging from 17.58 to 34.05 µM. SPR analysis displayed binding affinity of these molecules ranging from 8.13E-06 to 7.69E-04 for both the proteins. In the mouse bioassay, the selected molecules displayed complete protection and extension in survival of up to 20-fold. MD simulation study supported the experimental finding, revealing key interactions with HExxH and other active site residues forming stable conformation throughout the simulation time. Among them, compounds A15 and A36 were predicted to be more effective inhibitors of BoNT/B and F serotypes, respectively. These findings could lay a promising way for the development of novel therapeutics by reducing disease severity, enhancing survivability, and recovery where no post-exposure therapy presently available.
2026-01-17 | Therapeutic potential of [(8-hydroxyquinolin-7-yl)(phenyl)methylamino] benzoic acid regioisomers against human-intoxicating botulinum neurotoxin serotypes: computational modeling to in vivo protection.
Botulinum neurotoxins are class I tier bioterrorism agent, accountable for causing rare but fatal illness ‘botulism’. Out of seven serotypes, A, B, E, and F are responsible for intoxicating humans. Despite knowing harmful effects on human health for centuries, there is no commercial antidote for post-neuronal intoxication is available. In the present study, we report efficacy of regioisomers of [(8-hydroxyquinolin-7-yl)(phenyl) methylamino]benzoic acid against zinc-dependent light chain activities of BoNT/A, /B, /E & /F by combining molecular modeling with in vitro and in vivo studies. Based on structure similarity search, multiple regioisomers of 8-hydroxyquinoline were mined and screened by performing molecular docking. The best-scored compounds were analyzed for inhibitory and binding potential against these serotypes via endopeptidase and surface plasmon resonance assays. The best two compounds (NSC1011 and NSC1012) with potential inhibition and binding kinetics across serotypes were evaluated for therapeutic potential in mouse model. NSC1011 and NSC1012 (regioisomers) docking data revealed their binding energies with active domains of BoNT/A, /B, /E, and /F light chains ranging between − 9.70 to − 4.27, and − 9.84 to − 7.23 kcal/mol, respectively. The endopeptidase assay displayed ˃ 90% inhibition of catalytic activities, with the IC50 values varying among serotypes from 20 to 40 µM concentrations. SPR interaction of both compounds with the targeted proteins was observed in the range of 3.83E-05 to 4.95E-04 M. These molecules have shown complete protection at one MLD (mouse lethal dose), whereas median extension of animal survival was recorded up to 24 h when exposed to 5X MLD. The in silico, in vitro, and in vivo data reveal that NSC1011 and NSC1012 exhibited good binding affinity, stability, inhibition with promising therapeutic potential against human botulism-causing toxinotypes.
2025-11-12 | Two Cases of Severe Food-Borne Botulism: Could High-Dose Methylcobalamin Accelerate Recovery?
Botulinum neurotoxins (BoNTs) are zinc-dependent endopeptidases that cleave presynaptic SNARE proteins and arrest acetylcholine release, producing flaccid paralysis with protracted recovery. We report two cases of a mother and son who developed severe food-borne botulism after eating Fesikh (traditional salted fish) with rapidly progressive descending paralysis requiring ventilation, prominent dysautonomia (hypotension responsive to fluids, bradycardia, dry mucosae, constipation with abdominal distension, as well as urinary retention), and intact consciousness. Both received trivalent antitoxin on day 2; the son was extubated on day 10, and the mother on day 16. Persistent bladder atony, fatigue, weakness, and poor concentration improved with structured physiotherapy accompanied by intramuscular methylcobalamin, which was initiated on day 1 of admission and subsequently transitioned to an eight-week course of oral methylcobalamin (500 µg daily) during the post-extubation rehabilitation phase. We discuss mechanistic plausibility and hypothesize that higher-dose methylcobalamin might accelerate neuromuscular recovery by stabilizing mitochondria, attenuating caspase-mediated apoptosis, and promoting axonal regeneration and sprouting.
2025-10-14 | Phosphorylation of SNAP-25 at Ser187 is enhanced following its cleavage by Botulinum Neurotoxin Serotype A, promoting the dominant-negative effect of the resulting fragment.
Botulinum Neurotoxin Serotype A (BoNT/A), responsible for most human botulism cases, inhibits neurotransmitter release by cleaving the target protein SNAP-25. Previous literature demonstrated that BoNT/A mediated cleavage of a small subset of the SNAP-25 pool, resulting in SNAP-25 (1-197) fragments, is sufficient to block exocytosis. SNAP-25 (1-197) potentially competes against intact SNAP-25 for SNARE complexes and blocks neurotransmission through a dominant-negative mechanism. However, how a tiny fraction of cleaved SNAP-25 efficiently outcompetes a large pool of intact SNAP-25 remains unknown. Here, we examined the importance of SNAP-25 phosphorylation at Ser187 residue, located in the C-terminus SNARE domain, in the context of BoNT action. Our results demonstrated that Ser187-phosphorylated SNAP-25 can be efficiently cleaved in cells. Importantly, BoNT/A-cleaved SNAP-25 fragments in neuronal and non-neuronal cells are heavily phosphorylated at Ser187 and localized on the cell membrane. SNAP-25 (1-197) binds to syntaxin-1A, and the interaction is enhanced by Ser187 phosphorylation. We also found that SNAP-25 (1-197) survives longer than the BoNT/A enzymatic component itself in cells. Molecular modeling suggested that SNAP-25 (1-197), phosphorylated or not, forms stable SNARE complexes; however, Ser187 phosphorylation induces local changes in surface electrostatic potential and dynamics of the complex. This study characterizes the molecular mechanism underlying the dominant-negative effect of SNAP-25 (1-197) on neurotransmission. This research could have implications for the future development of BoNT/A inhibitors and the generation of new BoNT/A clinical formulations by regulating the abundance of Ser187 phosphorylation in cleaved SNAP-25 fragments.
proteins
2026-03-06 | Investigating the dual nature of Clostridium botulinum: pathogenic mechanisms and therapeutic potentials.
Clostridium botulinum (C. botulinum) is known for producing one of the strongest neurotoxins, namely botulinum neurotoxin (BoNT). This toxin is responsible for causing botulism, which is considered a severe neurological disease. This comprehensive narrative review highlights the dual nature of C. botulinum as both a serious health threat and a groundbreaking therapeutic agent. Methods: A comprehensive literature search was conducted in Google Scholar, PubMed/Medline, and Scopus (1971-2025, 2020-2025 literary focus) using keywords related to botulism pathogenesis, clinical features, and therapeutic approaches. Key findings indicate that, despite advancements in botulism management and reduced mortality due to heptavalent botulinum antitoxin (HBAT) and botulism immune globulin intravenous (BIG-IV) antitoxins, diagnostic challenges and bacterial spore resistance, particularly in group I, remain persistent. On the other hand, the discovery of serotype BoNT/X with lower toxicity in vertebrates and unique capabilities has opened new horizons in the design of safer treatments. C. botulinum represents a remarkable paradox in biology, an organism capable of producing one of the deadliest toxins known, yet simultaneously a cornerstone in modern therapeutic innovation. In fact, this toxin's dual nature is based on its selective targeting of BoNTs. In this regard, the systemic entry and distribution of this poison in the body - or, in other words, intoxication with it - poses a life-threatening aspect. On the other hand, the use of this bacterium's toxin for therapeutic and medicinal purposes is progressing due to its topical and controlled administration and entry into the body make it a targeted and effective therapeutic tool in the field of medicine. A better and more precise understanding of the mechanisms of action and, if necessary, future structural and engineered modifications in the toxin could lead to the emergence of a new generation of BoNTs that hold promise for the development of esthetic and therapeutic applications in medicine.
2026-01-27 | Advances in Clostridial Neurotoxins: Passage of the Intestinal Barrier and Targeting of Specific Neuronal Cells.
Clostridial neurotoxins, botulinum neurotoxins (BoNTs), and tetanus neurotoxin (TeNT) are potent toxins responsible for severe diseases, botulism and tetanus, respectively. BoNTs associate with non-toxic proteins (non-toxic non-hemagglutinin, hemagglutinins, and OrfXs), which protect BoNTs against acidic pH and protease degradation and facilitate BoNT passage through the intestinal barrier. TeNT enters motor neurons and undergoes a retrograde axonal transport until the target inhibitory interneurons in the central nervous system. BoNTs and TeNT recognize specific cell surface receptors which consist of complex sets of protein(s)-glycan-gangliosides and determine specific cell entry pathways. Recent data on structural and functional investigations of BoNT and TeNT receptors bring a better understanding of toxin trafficking in the host and entry into target neuronal cells, which is useful for the development of updated strategies of prevention and treatment of the corresponding diseases. Since clostridial neurotoxins, notably BoNTs, are important therapeutic tools, detailed knowledge of their activity opens the way of the development of engineered molecules for specific clinical applications.
2025-06-05 | Botulinum neurotoxin A mutants with enhanced ganglioside binding show improved potency and altered ganglioside selectivity.
Botulinum neurotoxins are the causative agents of botulism, a lethal paralytic disease, but are also one of the most commonly used therapeutics for the treatment of numerous neuromuscular conditions. These toxins recognise motor nerve terminals with high specificity and affinity by using a dual binding mechanism involving gangliosides and protein receptors. The initial recognition of gangliosides is crucial for the toxins' potency. In this study, we employed a synaptosome-binding screening strategy to identify BoNT/A mutants with enhanced ganglioside-binding which translated into improved potency. X-ray crystallography and receptor-binding assays were used to elucidate the molecular mechanisms underlying the increased affinity or altered ganglioside selectivity of these mutants. Our findings provide a basis for the development of BoNT/A variants with enhanced therapeutic potential.
2025-04-23 | Recombinant Detoxified Holotoxin as a Potent Candidate Vaccine Against Botulism.
Vaccination may represent a suitable strategy for preventing botulism. The recombinant expression of toxin functional domains can induce effective immune responses against botulism. This study aims to develop a safe and effective recombinant detoxified full-length BoNTA vaccine. In this study, we engineered and mutated the toxin activity-related sites on the basis of the full-length BoNTA protein and constructed three detoxified full-length toxin mutants. They were recombinant expressed and purified in Escherichia coli, and the BoNTA/M4 mutant was determined to have the highest safety, with a murine lethal dose of 50% (MLD50). The M4 protein was used as the antigen for three immunizations, and the serum titers, neutralizing activity, and BoNTA protective effects of immunized mice were evaluated. The results show that, in comparison to the receptor-binding domain BoNTA/Hc protein, the full-length detoxified mutant M4 protein exhibited superior immunogenicity and could induce higher levels of specific antibodies, and the resulting immune serum could effectively protect mice against higher doses of BoNTA challenge. This study laid the foundation for research on a novel recombinant detoxified full-length botulinum toxin vaccine.
2025-01-24 | Development of a Recombinant Fusion Vaccine Candidate Against Lethal Clostridium botulinum Neurotoxin Types A and B.
Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are potent protein toxins that can cause botulism, which leads to death or neuroparalysis in humans by targeting the nervous system. BoNTs comprise three functional domains: a light-chain enzymatic domain (LC), a heavy-chain translocation domain (HCN), and a heavy-chain receptor-binding domain (HCC). The HCC domain is critical for binding to neuronal cell membrane receptors and facilitating BoNT internalization via endocytosis. Accordingly, it may serve as a vaccine candidate, inducing anti-BoNT-neutralizing antibodies in animals. Here, we aimed to develop a vaccine capable of simultaneously defending against both BoNT/A and B. We combined the HCC domains of botulinum neurotoxin type A (BoNT/A) and botulinum neurotoxin type B (BoNT/B) in Escherichia coli to produce a recombinant protein (rHCCB-L-HCCArHCcB) that offers dual protection against both toxins by inhibiting their receptor binding. To evaluate the efficacy of the vaccine, mice were immunized intramuscularly with rHCCB-L-HCCA plus alum thrice at 2-week intervals, followed by the assessment of immunogenicity and protective efficacy. The antibody titer in mice immunized with rHCCB-L-HCCA was significantly higher than that in mice immunized with alum alone, protecting them from the lethal challenges of BoNT/A (105 50% lethal dose, LD50) and B (103 LD50). These findings suggest that rHCCB-L-HCCA may simultaneously be an effective vaccine candidate against BoNT/A and B.
antibodies
2026-06-17 | When BAT Precedes BIG-IV in Infant Botulism: The Need for Structured Antitoxin Decision-Pathway Reporting in Case Reports.
In infant botulism, use of equine-derived botulinum antitoxin before BIG-IV may be clinically justified when the infant-specific product is delayed. Future case reports should document the treatment decision pathway, antitoxin timing, risk-benefit justification, guardian counseling, adverse-event monitoring, and relapse-prevention rationale to make such emergency decisions reproducible.
2025-12-12 | Infant Botulism: A Case Study in Integrated Clinical and Public Health Response.
Early recognition of constipation, feeding difficulty, and descending weakness in infants is vital for prompt diagnosis of infant botulism. Bedside nerve studies support early treatment. Rapid public health notification and coordinated access to specific therapies, including timely use of BAT and BIG-IV, are essential to optimize outcomes and safety.
2025-12-12 | Whole-genome sequencing-based typing methods for Clostridium butyricum strains from clinical, animal, plant, and environmental sources.
Clostridium butyricum exhibits a dual role, acting not only as a probiotic but also as an opportunistic pathogen associated with neonatal necrotizing enterocolitis (NEC) and infant botulism. We aimed to establish high-resolution genotyping frameworks to improve molecular surveillance and outbreak investigations. We analyzed 297 C. butyricum genomes, including 200 isolates from preterm neonates across 13 French neonatal intensive care units over a 20-year period and 97 publicly available genomes. A core-genome multilocus sequence typing (cgMLST) scheme was developed using chewBBACA, defining 2,621 loci, and applied to genomes with ≥95% locus presence. Core-genome single-nucleotide polymorphism (cgSNP) analysis was performed for complementary resolution. Phylogenetic cgMLST classified isolates into nine major clades. Some clinical strains displayed clonal relationships, whereas others were geographically and temporally unrelated. All botulinum neurotoxin type E-producing strains were grouped within a single clade. NEC-associated isolates showed geographic and temporal clustering, but no clade was uniquely linked to NEC. cgSNP analysis identified 11 clusters with overall discriminatory power similar to cgMLST while providing finer resolution for NEC-related strains. We propose robust cgMLST and cgSNP schemes for C. butyricum, enabling high-resolution genotyping and supporting epidemiological surveillance and outbreak investigation of this emerging opportunistic pathogen in neonatal settings. Clostridium butyricum has been identified in fecal samples from both asymptomatic neonates and cases of necrotizing enterocolitis (NEC). Using a large collection of strains from different origins and spatiotemporal contexts, we developed and established a cgMLST scheme for the molecular typing of C. butyricum. Our results show that most C. butyricum strains cluster independently of origin and spatiotemporal context factors. However, specific cgMLST clades of C. butyricum were found for plant and botulinum neurotoxin type E strains. Clonal strains were also identified. No specific cgMLST clade was found to be genetically associated with NEC. cgSNP showed higher discriminatory power compared to cgMLST. Importantly, cgSNP provided better discriminatory power for strain relatedness with respect to strains isolated from NEC patients.
2025-11-26 | Regional Risk Discrepancy Indicates a Slow Outbreak of Infant Botulism, Victoria, Australia.
Infant botulism accounts for many new botulism cases each year. Geographic case clustering has been previously described in California. This study aimed to describe whether case clustering was present in Victoria, Australia. We conducted a retrospective case series of infant botulism presenting to Victorian hospitals between 1978 and 2024. Moran's I test for spatial autocorrelation was used to determine if spatiotemporal clustering was present. Twelve cases of infant botulism presented to Victorian hospitals in the study period. Using Moran's I statistic in analyzing the number of cases in each postcode of interest, a case clustering was demonstrated in the northwest of Victoria ( P < 0.01). We describe a geographic case clustering within the relatively sparsely populated northwest in Victoria, Australia.
2025-09-26 | Features of the First Case of Foodborne Botulism Caused by Dual-Toxin Clostridium parabotulinum Subtype A1(B5) in Spain.
The neurotoxin BoNT/B2 is the predominant Clostridium parabotulinum subtype in foodborne and infant botulism cases in Spain. This study characterizes a novel case of foodborne botulism in Spain caused by a dual-toxin A1(B5) strain. A 64-year-old male presented with acute, progressive flaccid paralysis including diplopia, dysphagia, and respiratory failure. Although botulism was not initially suspected, the patient recovered with supportive care and without antitoxin administration. Genomic characterization confirmed the presence of both bont/A1 and silent bont/B5 genes. The bont/A1 gene was associated with an orfX+ neurotoxin gene cluster, while the silent bont/B5 gene was in an ha+ cluster. Phylogenetic analysis of both bont/A1 and bont/B5 sequences showed 100% amino acid identity, respectively, to previously reported A1(B5) strains (e.g., CDC_69094, FE9504ACG). Multi-locus sequence typing (MLST) assigned the ST10, a genotype previously undetected in Spanish botulism cases, yet found in other European countries. This case highlights the importance of considering botulism in differential diagnosis due to its varied presentation and the significance of timely laboratory confirmation for effective management. The identification of this dual-toxin BoNT/A1(B5) orfX+/ha+ ST10 strain expands our understanding of C. botulinum epidemiology and genetic diversity in Spain.
vaccines
2026-01-27 | Proteolytic Activities and Immunological Effects of Light Chains of Botulinum Neurotoxin A1, A2 and A3 Subtypes.
Botulinum neurotoxin serotype A (BoNT/A) is the most potent known neurotoxin. While its light chain (LC) catalytic domain is a prime target for next-generation vaccines and therapeutics, the functional differences among BoNT/A subtype LCs (A1, A2, A3) remain to be definitively characterized, despite notable sequence variation. This work aimed to systematically compare the proteolytic activity and immunoprotective efficacy of recombinant BoNT/A1-LC, A2-LC, and A3-LC. Recombinant A1-LC-His, A2-LC-His, A3-LC-His, and A3-LC-Twin-Strep proteins were expressed in Escherichia coli (E. coli) and purified with affinity chromatography. Their proteolytic activity was assessed via in vitro SNAP-25 cleavage assays. The protective potency of these antigens was evaluated in a mouse model. In vitro cleavage assays revealed a substrate cleavage efficiency order of A2-LC > A1-LC > A3-LC. In vivo, both A1-LC and A2-LC immunization conferred robust, broad protection against high-dose challenges with all three toxin subtypes. In stark contrast, A3-LC provided only minimal protection against its homologous toxin and none against heterologous subtypes. Crucially, the functional deficit of A3-LC was confirmed to be an intrinsic property, as the A3-LC-TS variant, designed to exclude tag-specific interference, exhibited comparable low efficacy. According to structural research, A3-LC's compromised function may be caused by a four-amino-acid loss. The inferior performance of A3-LC is inherent to its primary structure. This work identified A1-LC or A2-LC as the potential proteolytic activity molecule and vaccine antigen by demonstrating functional differences among BoNT/A subtype LCs. These findings provide crucial insights for developing subtype-specific countermeasures against botulism.
2025-12-16 | Self-Applicable Thermostable Heterogeneous Microneedle Patch for Recombinant Protein Vaccination against Botulinum Neurotoxin Serotype A.
Botulinum neurotoxin serotype A (BoNT/A), a Class A bioterrorism agent, poses significant poisoning risks, with diagnosis challenges and no specific antidotes, making vaccination the most effective prevention. However, traditional needle-based vaccines are painful, thermally unstable, require trained professionals for administration, and carry infection risks, leading to low patient compliance. To overcome these limitations, we introduce a self-applicable, thermostable, heterogeneous microneedle (MN) patch for delivering a recombinant protein vaccine based on the C-terminal heavy chain of BoNT/A to prevent botulism. By incorporating materials with varying hydrophilicity, the MN patch enhances drug-layer contact with skin, improving vaccine delivery efficiency. Fabricated via heat- and pressure-assisted micromolding, the polycaprolactone MNs demonstrate sufficient mechanical strength for effective skin penetration. In vivo studies in a mouse model show that MN patches loaded with the recombinant C-terminal heavy chain of BoNT/A elicit strong IgG antibody responses and provide protective efficacy comparable to those of intramuscular injections, with all immunized mice surviving lethal BoNT/A challenges. The IgG responses of MN patches did not decrease after storage for 6 mol (25 °C, 60% relative humidity), indicating good stability. These findings suggest that thermostable MN patches offer a noninvasive, effective, patient-friendly alternative to traditional vaccines, with potential applications in bioterrorism prevention.
2025-04-18 | Detoxification techniques for bacterial toxins: A pathway to effective toxoid vaccines.
Bacterial toxins play a critical role in the virulence of many pathogens, leading to serious diseases such as tetanus, diphtheria, botulism, and entrotoxemia. As key virulence factors, these toxins cause significant tissue damage and disease manifestations in infected hosts. Vaccination against these toxins through toxoid vaccines, composed of inactivated forms of the toxins, represents a vital strategy for preventing toxin-mediated diseases. However, creating effective toxoid vaccines necessitates meticulous detoxification processes that ensure the loss of toxicity while retaining the immunogenic properties inherent in the native toxins. This review offers a comprehensive evaluation of the diverse methodologies employed for detoxifying bacterial toxins, highlighting their advantages, limitations, and implications for vaccine development. By detailing comparisons of efficacy, stability, residual toxicity, and clinical applicability, we demonstrate that while traditional methods utilizing chemical reagents (such as formaldehyde) remain widely used, emerging technologies like genetic inactivation and protein engineering present significant advantages. These innovations promise to advance the development of durable and irreversible toxoid vaccines that protect public health and contribute to future vaccine formulation improvements. Ultimately, this knowledge synthesis aims to guide future research efforts and facilitate the design of safer and more effective toxoid vaccines to combat the public health threats posed by toxin-producing bacteria.
2024-10-02 | New Engineered-Chimeric Botulinum Neurotoxin Mutant Acts as an Effective Bivalent Vaccine Against Botulinum Neurotoxin Serotype A and E.
Botulinum neurotoxins (BoNTs), including serotypes A and E, are potent biotoxins known to cause human poisoning. In addition to the critical protective antigen found in the full BoNT molecule, the receptor binding domain (Hc domain), BoNTs also harbour another essential protective antigen-the light chain-translocation domain (L-HN domain). Leveraging these pivotal protective antigens, we genetically engineered a series of inactivated chimeric molecules incorporating L-HN and Hc domains of BoNT/A and E. The structure of these chimeric molecules, mirror BoNT/A and E, but are devoid of enzyme activity. Experimental findings demonstrated that a lead candidate mEL-HN-mAHc harnessing the inactivated protease LCHN/E with the mutated gangliosides binding site Hc/A (mE-mA) elicited robust immune protection against BoNT/A and E simultaneously in a mouse model, requiring low immune dosages and minimal immunisations. Moreover, mE-mA exhibited high protective efficacy against BoNT/A and E in guinea pigs and New Zealand white rabbits, resulting in elevated neutralising antibody titres. Furthermore, mE-mA proved to be a more stable and safer vaccine compared to formaldehyde-inactivated toxoid. Our data underscore the genetically engineered mE-mA as a highly effective bivalent vaccine against BoNT/A and E, paving the way for the development of polyvalent vaccines against biotoxins.
2024-09-24 | Novel platform for engineering stable and effective vaccines against botulinum neurotoxins A, B and E.
Botulinum neurotoxin (BoNT), produced by Clostridium botulinum, is the most toxic protein known, capable of causing severe paralysis and posing a significant bioterrorism threat due to its extreme lethality even in minute quantities. Despite this, there are currently no FDA-approved vaccines for widespread public use. To address this urgent need, we have developed an innovative vaccine platform by fusing the neuronal binding domain of BoNT/E (Hc/E) with core-streptavidin (CS), resulting in a stable CS-Hc/E vaccine. Mice vaccinated with CS-Hc/E exhibited superior antibody titers compared to those receiving Hc/E alone. To develop a trivalent vaccine against BoNT/A, BoNT/B, and BoNT/E- key contributors to the vast majority of human botulism-we conjugated CS-Hc/E with a biotinylated atoxic chimeric protein incorporating neutralizing epitopes from BoNT/A and BoNT/B. This chimeric protein includes the binding domain of BoNT/A, along with the protease-inactive light chain and translocation domains of BoNT/B. The interaction between CS and biotin formed a stable tetrameric antigen, EBA. Vaccination with EBA in mice elicited robust antibody responses and provided complete protection against lethal doses of BoNT/A, BoNT/B, and BoNT/E. Our findings highlight EBA's potential as a stable and effective broad-spectrum vaccine against BoNT. Moreover, our technology offers a versatile platform for developing multivalent, stable vaccines targeting various biological threats by substituting the BoNT domain(s) with neutralizing epitopes from other life-threatening pathogens, thereby enhancing public health preparedness and biodefense strategies.
other
2026-06-24 | Acquisition of Stickland-metabolizing bacteria during infancy prevents Clostridium botulinum infection
Summary The mechanisms by which maturation of the infant gut microbiota promotes resistance to pathogen colonization remain poorly understood. Infant botulism, a potentially fatal disease caused by intestinal colonization by Clostridium botulinum , provides a striking example of age-dependent susceptibility to infection 1 . Although the gut microbiota has long been implicated in protection against C. botulinum 2–6 , the responsible bacterial species and underlying mechanisms have yet to be elucidated. Here, we show that acquisition of Stickland-metabolizing Clostridia during infant gut microbiota maturation confers resistance to C. botulinum colonization through competition for shared amino-acid-dependent nutritional niches. In human fecal microbiota-transplanted mice, longitudinally collected infant microbiotas exhibited a clear transition from susceptibility to resistance. Intestinal metabolomic analysis identified 5-aminovalerate as a hallmark metabolite of the resistant microbiota, implicating Stickland metabolism, an amino acid metabolic pathway also utilized by C. botulinum . Guided by this finding, metagenomic analysis revealed enrichment of Stickland-metabolizing Clostridia in resistant microbiotas, including Clostridioides difficile , a bacterium frequently carried by healthy infants. Intestinal metabolic signatures of C. botulinum and infant-derived Stickland-metabolizing Clostridia suggested competition for shared amino-acid-dependent nutritional niches. Consistent with this model, C. difficile suppressed C. botulinum expansion through nutrient competition. Together, these findings identify nutritional niche competition as a mechanism by which microbiota maturation promotes resistance to C. botulinum colonization. This work demonstrates how acquisition of specific microbial metabolic functions during early life can shift the gut microbiota from a susceptible to a resistant state.
2026-02-12 | Endemic koloa maoli (Hawaiian Duck, Anas wyvilliana ) shows preferential social associations, but not based on plumage or genetic relatedness.
Island endemic birds are under greater threat than their mainland counterparts. Sedentary living and historically reduced predation may affect island bird sociality and inform their conservation and management. However, detailed studies on their sociality are uncommon. The federally- endangered koloa maoli ( Anas wyvilliana, Hawaiian duck, or koloa) is primarily threatened by hybridization with feral mallards and avian botulism outbreaks. We used capture-mark-recapture and genetic data on koloa on the island of Kaua'i, a stronghold of remaining koloa, to construct social networks and examine their associations (inferred from co-occurrence in traps) and spatial genetic structure. Information on associations might shed light on preferences for or against mallards and hybrids, and inform planned translocation efforts. Microgeographic spatial genetic structuring where relatedness among individuals scales with geographic distance is a potential liability for maintaining koloa genetic diversity, and would particularly be detrimental during highly localized outbreaks of botulism that could result in the removal of entire lineages. While we found persistent social associations among adult koloa, they were not apparently influenced by plumage traits or body mass, suggesting a lack of social preference for mallard-like individuals. Importantly, we found no spatial patterns of relatedness within the largest refuge. Therefore, botulism outbreaks remain a demographic threat but are unlikely to remove correlated genetic diversity. There were no sex differences in spatial genetic structure and both sexes moved within a refuge. The lack of spatial genetic structure and the presence of many unrelated conspecifics may enable koloa to limit inbreeding and retain genetic diversity without sex-biased dispersal. In the context of future translocations, our results suggest that translocating koloa captured in the same trap together will reduce disruption of preferred associations while also retaining genetic diversity among translocated individuals. The koloa maoli (Hawaiian duck, or koloa) is a federally-endangered, island endemic bird. Like other Hawaiian waterbirds, koloa are threatened by introduced predators and habitat loss, but also specifically by hybridization with feral mallards and localized avian botulism outbreaks. Currently, the island of Kaua'i has the largest population of non-admixed koloa. We used capture-mark-recapture and genetic data of koloa at two wetland sites on Kaua'i to examine their sociality and spatial genetic structure.Koloa formed preferential social associations, but they were not based on plumage traits, body mass or genetic relatedness.There was no spatial genetic structure for males and females within a wetland site. Our results suggested that 1) koloa have no preference for mallard-like plumage that might increase hybrid pairing, 2) localized (within-refuge) botulism outbreaks are unlikely to remove close relatives and unique genetic lineages, and 3) translocation of groups could maintain social associations without limiting genetic diversity.
2024-01-02 | Development and characterization of a novel neutralizing scFv vectored immunoprophylaxis against botulinum toxin type A.
Botulinum toxin is a protein toxin secreted by Clostridium botulinum that is strongly neurotoxic. Due to its characteristics of being super toxic, quick acting, and difficult to prevent, the currently reported antiviral studies focusing on monoclonal antibodies have limited effectiveness. Therefore, for the sake of effectively prevention and treatment of botulism and to maintain country biosecurity as well as the health of the population, in this study, we intend to establish a single chain antibody (scFv) targeting the carboxyl terminal binding functional domain of the botulinum neurotoxin heavy chain (BONT/AHc) of botulinum neurotoxin type A, and explore the value of a new passive immune method in antiviral research which based on adeno-associated virus (AAV) mediated vector immunoprophylaxis (VIP) strategy. The scFv small-molecular single-chain antibody sequenced, designed, constructed, expressed and purified by hybridoma has high neutralising activity and affinity level, which can lay a good foundation for the modification and development of antibody engineering drugs. In vivo experiments, AAV-mediated scFv engineering drug has good anti-BONT/A toxin neutralisation ability, has advantages of simple operation, stable expression and good efficacy, and may be one of the effective treatment strategies for long-term prevention and protection of BONT/A botulinum neurotoxin.
2022-10-30 | Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A
ABSTRACT Single-domain antibodies (sdAbs, VHHs, or nanobodies) are a promising tool for the treatment of both infectious and somatic diseases. Their small size greatly simplifies any genetic engineering manipulations. Such antibodies have the ability to bind hard-to-reach antigenic epitopes through long parts of the variable chains, the third complementarity-determining regions (CDR3s). VHH fusion with the canonical immunoglobulin Fc fragment allows the Fc-fusion single-domain antibodies (VHH-Fc) to significantly increase their neutralizing activity and serum half-life. Previously we have developed and characterized VHH-Fc specific to botulinum neurotoxin A (BoNT/A), that showed a 1000-fold higher protective activity than monomeric form when challenged with five times the lethal dose (5 LD 50 ) of BoNT/A. During the COVID-19 pandemic, mRNA vaccines based on lipid nanoparticles (LNP) as a delivery system have become an important translational technology that has significantly accelerated the clinical introduction of mRNA platforms. We have developed an mRNA platform that provides long-term expression after both intramuscular and intravenous application. The platform has been extensively characterized using firefly luciferase (Fluc) as a reporter. An intramuscular administration of LNP-mRNA encoding VHH-Fc antibody made it possible to achieve its rapid expression in mice and resulted in 100% protection when challenged with up to 100 LD 50 of BoNT/A. The presented approach for the delivery of sdAbs using mRNA technology greatly simplifies drug development for antibody therapy and can be used for emergency prophylaxis.
2022-06-30 | Genomic and Phenotypic Characterization of Clostridium botulinum Isolates from an Infant Botulism Case Suggests Adaptation Signatures to the Gut.
In early life, the immature human gut microbiota is prone to colonization by pathogens that are usually outcompeted by mature microbiota in the adult gut. Colonization and neurotoxin production by a vegetative Clostridium botulinum culture in the gut of an infant can lead to flaccid paralysis, resulting in a clinical outcome known as infant botulism, a potentially life-threatening condition. Beside host factors, little is known of the ecology, colonization, and adaptation of C. botulinum to the gut environment. In our previous report, an infant with intestinal botulism was shown to be colonized by neurotoxigenic C. botulinum culture for 7 months. In an effort to gain ecological and evolutionary insights into this unusually long gut colonization by C. botulinum, we analyzed and compared the genomes of C. botulinum isolates recovered from the infant feces during the course of intoxication and isolates from the infant household dust. A number of observed mutations and genomic alterations pinpointed at phenotypic traits that may have promoted colonization and adaptation to the gut environment and to the host. These traits include motility, quorum-sensing, sporulation, and carbohydrate metabolism. We provide novel perspectives and suggest a tentative model of the pathogenesis of C. botulinum in infant botulism. IMPORTANCE While the clinical aspects of infant botulism and the mode of action of BoNT have been thoroughly investigated, little is known on the pathogenesis and adaptive mechanisms of C. botulinum in the gut. Here, we provide for the first time a comprehensive view on the genomic dynamics and plasticity of C. botulinum over time in a case of infant botulism. The genomic and phenotypic analysis of C. botulinum isolates collected during the disease course offers an unprecedented view of C. botulinum ecology, evolution, and pathogenesis and may be instrumental in developing novel strategies for prevention and treatment of toxicoinfectious botulism.
Access all drug discovery papers and probability of success in trials forecasts:
Access all drug discovery papers and probability of success in trials forecasts:
Drug Discovery Landscape
1 orphan drug designation for Infant botulism, including 1 approved therapy.
1 orphan drug designation for Infant botulism, including 1 approved therapy.
Drug | Therapy type | Regulator | Orphan designation | Approval | Sponsor |
|---|---|---|---|---|---|
Botulism immune globulin [BabyBIG] | antibodies | FDA | 1989-01-31 | 2003-10-23 | California Department of Public Health |
Let's accelerate rare disease drug discovery
Let's accelerate drug discovery
Get access to Explority AI’s forecasts to outperform average preclinical success rates. Whether you’re expanding your R&D pipeline, evaluating a partnership, or simply have a question — we’d love to hear from you.