Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin

Garcia-Rodriguez, Consuelo; Lévy, Raphaël; Arndt, Joseph W.; Forsyth, Charles M.; Razai, Ali; Lou, Jianlong; Geren, Isin N.; Stevens, Raymond C.; Marks, James D.

doi:10.1038/nbt1269

Cited by 162 publications

(167 citation statements)

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“…The resulting refinements in existing contacts led to dramatic improvements in kDa and increased buried surface area between E10 and CXCL13, in comparison with the parental clone 3B4. Our findings echo those of Garcia-Rodriguez et al (29), who demonstrated that CDR-based mutations improving the affinity of a BoNT/A1-A2 cross-reactive antibody for the A2 form eliminated an electrostatic clash while also generating a new energetic interaction.…”

Section: Discussionsupporting

confidence: 52%

A Combination of Structural and Empirical Analyses Delineates the Key Contacts Mediating Stability and Affinity Increases in an Optimized Biotherapeutic Single-chain Fv (scFv)

Terraube

Tam

et al. 2016

Journal of Biological Chemistry

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Background: Antibody v-domains in scFv format often suffer from aggregation and stability issues that restrict formulation. Results: Structural and empirical analyses of an optimized scFv revealed that three V L -CDR3 mutations were sufficient to mediate significant stability and affinity improvements. Conclusion: scFv issues were resolved via removal of side-chain clashes at the V L /V H interface. Significance: CDR-restricted mutagenesis delivers stability-optimized molecules for high concentration dosing.

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Section: Discussionsupporting

confidence: 52%

A Combination of Structural and Empirical Analyses Delineates the Key Contacts Mediating Stability and Affinity Increases in an Optimized Biotherapeutic Single-chain Fv (scFv)

Terraube

Tam

et al. 2016

Journal of Biological Chemistry

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“…Collected serum were aliquoted and stored at Ϫ80°C before analysis. MSD assays used human MAb ING2 for toxin capture and SULFO-TAG-labeled MAb CR2 as a reporter (13,16). Assay protocols were optimized after screening and comparing eight different MAbs specific for BoNT/A.…”

Section: Methodsmentioning

confidence: 99%

Antibody Protection against Botulinum Neurotoxin Intoxication in Mice

et al. 2009

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Adulteration of food or feed with any of the seven serotypes of botulinum neurotoxin (BoNT) is a potential bioterrorism concern. Currently, there is strong interest in the development of detection reagents, vaccines, therapeutics, and other countermeasures. A sensitive immunoassay for detecting BoNT serotype A (BoNT/A), based on monoclonal antibodies (MAbs) F1-2 and F1-40, has been developed and used in complex matrices. The epitope for F1-2 has been mapped to the heavy chain of BoNT/A, and the epitope of F1-40 has been mapped to the light chain. The ability of these MAbs to provide therapeutic protection against BoNT/A intoxication in mouse intravenous and oral intoxication models was tested. High dosages of individual MAbs protected mice well both pre-and postexposure to BoNT/A holotoxin. A combination therapy consisting of antibodies against both the light and heavy chains of the toxin, however, significantly increased protection, even at a lower MAb dosage. An in vitro peptide assay for measuring toxin activity showed that pretreatment of toxin with these MAbs did not block catalytic activity but instead blocked toxin entry into primary and cultured neuronal cells. The timing of antibody rescue in the mouse intoxication models revealed windows of opportunity for antibody therapeutic treatment that correlated well with the biologic half-life of the toxin in the serum. Knowledge of BoNT intoxication and antibody clearance in these mouse models and understanding of the pharmacokinetics of BoNT are invaluable for future development of antibodies and therapeutics against intoxication by BoNT.Botulinum neurotoxins (BoNTs) are considered some of the most potent toxins known and are potential bioterrorist threat agents. Yet, BoNT serotype A (BoNT/A) and BoNT/B are also used therapeutically in a wide array of medical conditions, such as dystonia and eye disorders like strabismus and blepharospasms, for pain management, and more (3, 25). Thus, there is a need to protect humans and animals against toxin exposure from contaminated food or feed and yet preserve the medical benefits of BoNT. A better understanding of the biology of the toxin, such as toxin distribution and mechanisms of toxin neutralization following intoxication, is needed to aid further development of improved therapies, as well as bona fide use of toxin to treat serious medical conditions.BoNTs are 150-kDa endopeptidase toxins that are produced by Clostridium botulinum, C. butyricum, and C. baratii (20,26,27). The toxin polypeptide is cleaved upon secretion from the cell by bacterial proteases or proteases in the animal host into a disulfide bond-linked dipeptide consisting of a 100-kDa heavy chain (Hc) and a 50-kDa light chain (Lc). The 50-kDa Lc contains the active site or catalytic domain that targets the soluble N-ethylmaleimide-sensitive factor attachment protein receptor. Specifically, the synaptosome-associated 25-kDa protein (SNAP25) is the target for BoNT/A. Cleavage of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor ...

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“…Engineering specificity includes reducing the cross-reactivity to other antigens, broadening the specificity to related antigens, and improving the cross-reactivity with nonhuman species [292][293][294][295][296][297][298][299][300][301]. This can be achieved by the same mutagenesis and display technologies used for affinity maturation as phage display technology or computational design approach.…”

Section: Specificity Of Antibodiesmentioning

confidence: 99%

Applications of immunochemistry in human health: advances in vaccinology and antibody design (IUPAC Technical Report)

Klein

Templeton

Schwenk

2014

Pure and Applied Chemistry

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This report discusses the history and mechanisms of vaccination of humans as well as the engineering of therapeutic antibodies. Deeper understanding of the molecular interactions involved in both acquired and innate immunity is allowing sophistication in design of modified and even synthetic vaccines. Recombinant DNA technologies are facilitating development of DNA-based vaccines, for example, with the recognition that unmethylated CpG sequences in plasmid DNA will target Toll-like receptors on antigen-presenting cells. Formulations of DNA vaccines with increased immunogenicity include engineering into plasmids with "genetic adjuvant" capability, incorporation into polymeric or magnetic nanoparticles, and formulation with cationic polymers and other polymeric and non-polymeric coatings. Newer methods of delivery, such as particle bombardment, DNA tattooing, electroporation, and magnetic delivery, are also improving the effectiveness of DNA vaccines. RNA-based vaccines and reverse vaccinology based on gene sequencing and bioinformatic approaches are also considered. Structural vaccinology is an approach in which the detailed molecular structure of viral epitopes is used to design synthetic antigenic peptides. Virus-like particles are being designed for vaccine deliveries that are based on structures of viral capsid proteins and other synthetic lipopeptide building blocks. A new generation of adjuvants is being developed to further enhance immunogenicity, based on squalene and other oil-water emulsions, saponins, muramyl dipeptide, immunostimulatory oligonucleotides, Toll-like receptor ligands, and lymphotoxins. Finally, current trends in engineering of therapeutic antibodies including improvements of antigen-binding properties, pharmacokinetic and pharmaceutical properties, and reduction of immunogenicity are discussed. Taken together, understanding the chemistry of vaccine design, delivery and immunostimulation, and knowledge of the techniques of antibody design are allowing targeted development for the treatment of chronic disorders characterized by continuing activation of the immune system, such as autoimmune disorders, cancer, or allergies that have long been refractory to conventional approaches.

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Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin

Cited by 162 publications

References 46 publications

A Combination of Structural and Empirical Analyses Delineates the Key Contacts Mediating Stability and Affinity Increases in an Optimized Biotherapeutic Single-chain Fv (scFv)

A Combination of Structural and Empirical Analyses Delineates the Key Contacts Mediating Stability and Affinity Increases in an Optimized Biotherapeutic Single-chain Fv (scFv)

Antibody Protection against Botulinum Neurotoxin Intoxication in Mice

Applications of immunochemistry in human health: advances in vaccinology and antibody design (IUPAC Technical Report)

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