FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Mol Biol / Fine and domain-level epitope mapping of botulinum neurotoxi...
Journal of molecular biology2006; 365(1); 196-210; doi: 10.1016/j.jmb.2006.09.084

Fine and domain-level epitope mapping of botulinum neurotoxin type A neutralizing antibodies by yeast surface display.

Abstract: Botulinum neurotoxin (BoNT), the most poisonous substance known, causes naturally occurring human disease (botulism) and is one of the top six biothreat agents. Botulism is treated with polyclonal antibodies produced in horses that are associated with a high incidence of systemic reactions. Human monoclonal antibodies (mAbs) are under development as a safer therapy. Identifying neutralizing epitopes on BoNTs is an important step in generating neutralizing mAbs, and has implications for vaccine development. Here, we show that the three domains of BoNT serotype A (BoNT/A) can be displayed on the surface of yeast and used to epitope map six mAbs to the toxin domains they bind. The use of yeast obviates the need to express and purify each domain, and it should prove possible to display domains of other BoNT subtypes and serotypes for epitope mapping. Using a library of yeast-displayed BoNT/A binding domain (H(C)) mutants and selecting for loss of binding, the fine epitopes of three neutralizing BoNT/A mAbs were identified. Two mAbs bind the C-terminal subdomain of H(C), with one binding near the toxin sialoganglioside binding site. The most potently neutralizing mAb binds the N-terminal subdomain of H(C), in an area not previously thought to be functionally important. Modeling the epitopes shows how all three mAbs could bind BoNT/A simultaneously and may explain, in part, the dramatic synergy observed on in vivo toxin neutralization when these antibodies are combined. The results demonstrate how yeast display can be used for domain-level and fine mapping of conformational BoNT antibody epitopes and the mapping results identify three neutralizing BoNT/A epitopes.
Get Full Text
Publication Date: 2006-10-03 PubMed ID: 17059824PubMed Central: PMC1994578DOI: 10.1016/j.jmb.2006.09.084Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the process of identifying neutralizing epitopes on Botulinum neurotoxin type A using yeast surface display, with the aim of developing human monoclonal antibodies for safer treatment of botulism. The research also includes detailed epitope mapping and implications for vaccine development.

Objective of the Research

  • The aim of the study was to identify neutralizing epitopes on Botulinum neurotoxin type A (BoNT/A) using yeast surface display.
  • This is a crucial step in generating neutralizing monoclonal antibodies for the treatment of botulism, a severe condition caused by the most poisonous substance known – botulinum neurotoxin.
  • The researchers also intend to map the respective domains of the toxin the antibodies bind to, which can contribute significantly to vaccine development.

Methodology

  • The researchers displayed the three domains of BoNT/A on the surface of yeast for epitope mapping with six monoclonal antibodies.
  • This process of using yeast eliminates the need to express and purify each domain separately.
  • A library of yeast-displayed BoNT/A binding domain (H(C)) mutants was used and selected for loss of binding to identify the fine epitopes of three neutralizing antibodies.

Findings

  • Two monoclonal antibodies were found to bind the C-terminal subdomain of H(C), with one binding near the toxin sialoganglioside binding site.
  • The most potently neutralizing antibody was found to bind the N-terminal subdomain of H(C), an area not previously considered crucial.
  • Through modelling, the researchers demonstrated that all three antibodies could bind BoNT/A simultaneously, which could partly explain the significant synergy observed in in vivo toxin neutralization when these antibodies are combined.

Significance and Conclusion

  • The study indicates that yeast display can be effectively used for domain-level and fine mapping of conformational BoNT antibody epitopes, providing key insights into the neutralizing mechanisms.
  • The mapping results identified three neutralizing BoNT/A epitopes, which could aid in the design of vaccines and contribute to the development of safer therapy options for botulism using human monoclonal antibodies.

Cite This Article

APA
Levy R, Forsyth CM, LaPorte SL, Geren IN, Smith LA, Marks JD. (2006). Fine and domain-level epitope mapping of botulinum neurotoxin type A neutralizing antibodies by yeast surface display. J Mol Biol, 365(1), 196-210. https://doi.org/10.1016/j.jmb.2006.09.084

Publication

Journal of molecular biology
ISSN: 0022-2836
NlmUniqueID: 2985088R
Country: Netherlands
Language: English
Volume: 365
Issue: 1
Pages: 196-210

Researcher Affiliations

Levy, R
  • Department of Anesthesia and Pharmaceutical Chemistry, University of California, San Francisco, San Francisco General Hospital, CA 94110, USA.
Forsyth, C M
    LaPorte, S L
      Geren, I N
        Smith, L A
          Marks, J D

            MeSH Terms

            • Antibodies, Bacterial / immunology
            • Antibodies, Monoclonal / immunology
            • Antibody Specificity
            • Antigen-Antibody Complex
            • Botulinum Toxins, Type A / chemistry
            • Botulinum Toxins, Type A / immunology
            • Clostridium botulinum / immunology
            • Epitope Mapping
            • Humans
            • Models, Molecular
            • Mutagenesis
            • Neutralization Tests
            • Peptide Library
            • Protein Structure, Tertiary
            • Saccharomyces cerevisiae

            Grant Funding

            • U01 AI056493 / NIAID NIH HHS

            References

            This article includes 48 references
            1. Gill DM. Bacterial toxins: a table of lethal amounts.. Microbiol Rev 1982 Mar;46(1):86-94.
              pmc: PMC373212pubmed: 6806598doi: 10.1128/mr.46.1.86-94.1982google scholar: lookup
            2. Lacy DB, Tepp W, Cohen AC, DasGupta BR, Stevens RC. Crystal structure of botulinum neurotoxin type A and implications for toxicity.. Nat Struct Biol 1998 Oct;5(10):898-902.
              pubmed: 9783750doi: 10.1038/2338google scholar: lookup
            3. Simpson LL. Kinetic studies on the interaction between botulinum toxin type A and the cholinergic neuromuscular junction.. J Pharmacol Exp Ther 1980 Jan;212(1):16-21.
              pubmed: 6243359
            4. Montecucco C, Schiavo G. Structure and function of tetanus and botulinum neurotoxins.. Q Rev Biophys 1995 Nov;28(4):423-72.
              pubmed: 8771234doi: 10.1017/s0033583500003292google scholar: lookup
            5. Dong M, Yeh F, Tepp WH, Dean C, Johnson EA, Janz R, Chapman ER. SV2 is the protein receptor for botulinum neurotoxin A.. Science 2006 Apr 28;312(5773):592-6.
              pubmed: 16543415doi: 10.1126/science.1123654google scholar: lookup
            6. Dolly JO, Black J, Williams RS, Melling J. Acceptors for botulinum neurotoxin reside on motor nerve terminals and mediate its internalization.. Nature 1984 Feb 2-8;307(5950):457-60.
              pubmed: 6694738doi: 10.1038/307457a0google scholar: lookup
            7. Mahrhold S, Rummel A, Bigalke H, Davletov B, Binz T. The synaptic vesicle protein 2C mediates the uptake of botulinum neurotoxin A into phrenic nerves.. FEBS Lett 2006 Apr 3;580(8):2011-4.
              pubmed: 16545378doi: 10.1016/j.febslet.2006.02.074google scholar: lookup
            8. Schiavo G, Benfenati F, Poulain B, Rossetto O, Polverino de Laureto P, DasGupta BR, Montecucco C. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin.. Nature 1992 Oct 29;359(6398):832-5.
              pubmed: 1331807doi: 10.1038/359832a0google scholar: lookup
            9. Schiavo G, Rossetto O, Catsicas S, Polverino de Laureto P, DasGupta BR, Benfenati F, Montecucco C. Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E.. J Biol Chem 1993 Nov 15;268(32):23784-7.
              pubmed: 8226912
            10. Centers for Disease Control. Handbook for Epidemiologists, Clinicians,and Laboratory Workers. Atlanta: Georgia U.S. Department of Health and Human Services, Public Health Service; 1998.
            11. Arnon SS, Schechter R, Inglesby TV, Henderson DA, Bartlett JG, Ascher MS, Eitzen E, Fine AD, Hauer J, Layton M, Lillibridge S, Osterholm MT, O'Toole T, Parker G, Perl TM, Russell PK, Swerdlow DL, Tonat K. Botulinum toxin as a biological weapon: medical and public health management.. JAMA 2001 Feb 28;285(8):1059-70.
              pubmed: 11209178doi: 10.1001/jama.285.8.1059google scholar: lookup
            12. Franz DR, Pitt LM, Clayton MA, Hanes MA, Rose KJ. In: Botulinum and Tetanus Neurotoxins: Neurotransmission and Biomedical Aspects. DasGupta BR, editor. Plenum Press; New York: 1993. pp. 473–476.
            13. Black RE, Gunn RA. Hypersensitivity reactions associated with botulinal antitoxin.. Am J Med 1980 Oct;69(4):567-70.
              pubmed: 7191633doi: 10.1016/0002-9343(80)90469-6google scholar: lookup
            14. Amersdorfer P, Wong C, Smith T, Chen S, Deshpande S, Sheridan R, Marks JD. Genetic and immunological comparison of anti-botulinum type A antibodies from immune and non-immune human phage libraries.. Vaccine 2002 Feb 22;20(11-12):1640-8.
              pubmed: 11858873doi: 10.1016/s0264-410x(01)00482-0google scholar: lookup
            15. Nowakowski A, Wang C, Powers DB, Amersdorfer P, Smith TJ, Montgomery VA, Sheridan R, Blake R, Smith LA, Marks JD. Potent neutralization of botulinum neurotoxin by recombinant oligoclonal antibody.. Proc Natl Acad Sci U S A 2002 Aug 20;99(17):11346-50.
              pmc: PMC123259pubmed: 12177434doi: 10.1073/pnas.172229899google scholar: lookup
            16. Swaminathan S, Eswaramoorthy S. Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B.. Nat Struct Biol 2000 Aug;7(8):693-9.
              pubmed: 10932256doi: 10.1038/78005google scholar: lookup
            17. Eswaramoorthy S, Kumaran D, Swaminathan S. Crystallographic evidence for doxorubicin binding to the receptor-binding site in Clostridium botulinum neurotoxin B.. Acta Crystallogr D Biol Crystallogr 2001 Nov;57(Pt 11):1743-6.
              pubmed: 11679763doi: 10.1107/s0907444901013531google scholar: lookup
            18. Boder ET, Wittrup KD. Yeast surface display for screening combinatorial polypeptide libraries.. Nat Biotechnol 1997 Jun;15(6):553-7.
              pubmed: 9181578doi: 10.1038/nbt0697-553google scholar: lookup
            19. Chao G, Cochran JR, Wittrup KD. Fine epitope mapping of anti-epidermal growth factor receptor antibodies through random mutagenesis and yeast surface display.. J Mol Biol 2004 Sep 10;342(2):539-50.
              pubmed: 15327953doi: 10.1016/j.jmb.2004.07.053google scholar: lookup
            20. Razai A, Garcia-Rodriguez C, Lou J, Geren IN, Forsyth CM, Robles Y, Tsai R, Smith TJ, Smith LA, Siegel RW, Feldhaus M, Marks JD. Molecular evolution of antibody affinity for sensitive detection of botulinum neurotoxin type A.. J Mol Biol 2005 Aug 5;351(1):158-69.
              pubmed: 16002090doi: 10.1016/j.jmb.2005.06.003google scholar: lookup
            21. Mullaney BP, Pallavicini MG, Marks JD. Epitope mapping of neutralizing botulinum neurotoxin A antibodies by phage display.. Infect Immun 2001 Oct;69(10):6511-4.
              pmc: PMC98787pubmed: 11553596doi: 10.1128/iai.69.10.6511-6514.2001google scholar: lookup
            22. Amersdorfer P, Wong C, Chen S, Smith T, Deshpande S, Sheridan R, Finnern R, Marks JD. Molecular characterization of murine humoral immune response to botulinum neurotoxin type A binding domain as assessed by using phage antibody libraries.. Infect Immun 1997 Sep;65(9):3743-52.
              pmc: PMC175534pubmed: 9284147doi: 10.1128/iai.65.9.3743-3752.1997google scholar: lookup
            23. Smith TJ, Lou J, Geren IN, Forsyth CM, Tsai R, Laporte SL, Tepp WH, Bradshaw M, Johnson EA, Smith LA, Marks JD. Sequence variation within botulinum neurotoxin serotypes impacts antibody binding and neutralization.. Infect Immun 2005 Sep;73(9):5450-7.
              pmc: PMC1231122pubmed: 16113261doi: 10.1128/iai.73.9.5450-5457.2005google scholar: lookup
            24. Fromant M, Blanquet S, Plateau P. Direct random mutagenesis of gene-sized DNA fragments using polymerase chain reaction.. Anal Biochem 1995 Jan 1;224(1):347-53.
              pubmed: 7710092doi: 10.1006/abio.1995.1050google scholar: lookup
            25. Cunningham BC, Wells JA. High-resolution epitope mapping of hGH-receptor interactions by alanine-scanning mutagenesis.. Science 1989 Jun 2;244(4908):1081-5.
              pubmed: 2471267doi: 10.1126/science.2471267google scholar: lookup
            26. Wells JA. Systematic mutational analyses of protein-protein interfaces.. Methods Enzymol 1991;202:390-411.
              pubmed: 1723781doi: 10.1016/0076-6879(91)02020-agoogle scholar: lookup
            27. Nielsen UB, Adams GP, Weiner LM, Marks JD. Targeting of bivalent anti-ErbB2 diabody antibody fragments to tumor cells is independent of the intrinsic antibody affinity.. Cancer Res 2000 Nov 15;60(22):6434-40.
              pubmed: 11103810
            28. Lacy DB, Stevens RC. Sequence homology and structural analysis of the clostridial neurotoxins.. J Mol Biol 1999 Sep 3;291(5):1091-104.
              pubmed: 10518945doi: 10.1006/jmbi.1999.2945google scholar: lookup
            29. Rummel A, Mahrhold S, Bigalke H, Binz T. The HCC-domain of botulinum neurotoxins A and B exhibits a singular ganglioside binding site displaying serotype specific carbohydrate interaction.. Mol Microbiol 2004 Feb;51(3):631-43.
              pubmed: 14731268doi: 10.1046/j.1365-2958.2003.03872.xgoogle scholar: lookup
            30. Tavallaie M, Chenal A, Gillet D, Pereira Y, Manich M, Gibert M, Raffestin S, Popoff MR, Marvaud JC. Interaction between the two subdomains of the C-terminal part of the botulinum neurotoxin A is essential for the generation of protective antibodies.. FEBS Lett 2004 Aug 13;572(1-3):299-306.
              pubmed: 15304366doi: 10.1016/j.febslet.2004.06.094google scholar: lookup
            31. Herreros J, Lalli G, Montecucco C, Schiavo G. Tetanus toxin fragment C binds to a protein present in neuronal cell lines and motoneurons.. J Neurochem 2000 May;74(5):1941-50.
              pubmed: 10800937doi: 10.1046/j.1471-4159.2000.0741941.xgoogle scholar: lookup
            32. Frank R, Overwin H. SPOT synthesis. Epitope analysis with arrays of synthetic peptides prepared on cellulose membranes.. Methods Mol Biol 1996;66:149-69.
              pubmed: 8959713doi: 10.1385/0-89603-375-9:149google scholar: lookup
            33. Christmann A, Wentzel A, Meyer C, Meyers G, Kolmar H. Epitope mapping and affinity purification of monospecific antibodies by Escherichia coli cell surface display of gene-derived random peptide libraries.. J Immunol Methods 2001 Nov 1;257(1-2):163-73.
              pubmed: 11687250doi: 10.1016/s0022-1759(01)00461-6google scholar: lookup
            34. Bénichou S, Inchauspé G. Random fragment libraries using yeast expression plasmid.. Methods Mol Biol 1996;66:241-55.
              pubmed: 8959720doi: 10.1385/0-89603-375-9:241google scholar: lookup
            35. Wang LF, Yu M. Epitope identification and discovery using phage display libraries: applications in vaccine development and diagnostics.. Curr Drug Targets 2004 Jan;5(1):1-15.
              pubmed: 14738215doi: 10.2174/1389450043490668google scholar: lookup
            36. Mehra V, Sweetser D, Young RA. Efficient mapping of protein antigenic determinants.. Proc Natl Acad Sci U S A 1986 Sep;83(18):7013-7.
              pmc: PMC386642pubmed: 2428046doi: 10.1073/pnas.83.18.7013google scholar: lookup
            37. Weiss GA, Watanabe CK, Zhong A, Goddard A, Sidhu SS. Rapid mapping of protein functional epitopes by combinatorial alanine scanning.. Proc Natl Acad Sci U S A 2000 Aug 1;97(16):8950-4.
              pmc: PMC16802pubmed: 10908667doi: 10.1073/pnas.160252097google scholar: lookup
            38. Vajdos FF, Adams CW, Breece TN, Presta LG, de Vos AM, Sidhu SS. Comprehensive functional maps of the antigen-binding site of an anti-ErbB2 antibody obtained with shotgun scanning mutagenesis.. J Mol Biol 2002 Jul 5;320(2):415-28.
              pubmed: 12079396doi: 10.1016/s0022-2836(02)00264-4google scholar: lookup
            39. Oliphant T, Engle M, Nybakken GE, Doane C, Johnson S, Huang L, Gorlatov S, Mehlhop E, Marri A, Chung KM, Ebel GD, Kramer LD, Fremont DH, Diamond MS. Development of a humanized monoclonal antibody with therapeutic potential against West Nile virus.. Nat Med 2005 May;11(5):522-30.
              pmc: PMC1458527pubmed: 15852016doi: 10.1038/nm1240google scholar: lookup
            40. Hall YH, Chaddock JA, Moulsdale HJ, Kirby ER, Alexander FC, Marks JD, Foster KA. Novel application of an in vitro technique to the detection and quantification of botulinum neurotoxin antibodies.. J Immunol Methods 2004 May;288(1-2):55-60.
              pubmed: 15183085doi: 10.1016/j.jim.2004.02.011google scholar: lookup
            41. Montero-Julian FA, Klein B, Gautherot E, Brailly H. Pharmacokinetic study of anti-interleukin-6 (IL-6) therapy with monoclonal antibodies: enhancement of IL-6 clearance by cocktails of anti-IL-6 antibodies.. Blood 1995 Feb 15;85(4):917-24.
              pubmed: 7849313
            42. Clayton MA, Clayton JM, Brown DR, Middlebrook JL. Protective vaccination with a recombinant fragment of Clostridium botulinum neurotoxin serotype A expressed from a synthetic gene in Escherichia coli.. Infect Immun 1995 Jul;63(7):2738-42.
              pmc: PMC173366pubmed: 7790092doi: 10.1128/iai.63.7.2738-2742.1995google scholar: lookup
            43. Potter KJ, Zhang W, Smith LA, Meagher MM. Production and purification of the heavy chain fragment C of botulinum neurotoxin, serotype A, expressed in the methylotrophic yeast Pichia pastoris.. Protein Expr Purif 2000 Aug;19(3):393-402.
              pubmed: 10910730doi: 10.1006/prep.2000.1256google scholar: lookup
            44. Smith LA, Jensen MJ, Montgomery VA, Brown DR, Ahmed SA, Smith TJ. Roads from vaccines to therapies.. Mov Disord 2004 Mar;19 Suppl 8:S48-52.
              pubmed: 15027054doi: 10.1002/mds.20009google scholar: lookup
            45. Ahmed SA, Smith LA. Light chain of botulinum A neurotoxin expressed as an inclusion body from a synthetic gene is catalytically and functionally active.. J Protein Chem 2000 Aug;19(6):475-87.
              pubmed: 11195972doi: 10.1023/a:1026549431380google scholar: lookup
            46. Jensen MJ, Smith TJ, Ahmed SA, Smith LA. Expression, purification, and efficacy of the type A botulinum neurotoxin catalytic domain fused to two translocation domain variants.. Toxicon 2003 May;41(6):691-701.
              pubmed: 12727273doi: 10.1016/s0041-0101(03)00042-4google scholar: lookup
            47. Gietz RD, Schiestl RH. Applications of high efficiency lithium acetate transformation of intact yeast cells using single-stranded nucleic acids as carrier.. Yeast 1991 Apr;7(3):253-63.
              pubmed: 1882550doi: 10.1002/yea.320070307google scholar: lookup
            48. Feldhaus MJ, Siegel RW, Opresko LK, Coleman JR, Feldhaus JM, Yeung YA, Cochran JR, Heinzelman P, Colby D, Swers J, Graff C, Wiley HS, Wittrup KD. Flow-cytometric isolation of human antibodies from a nonimmune Saccharomyces cerevisiae surface display library.. Nat Biotechnol 2003 Feb;21(2):163-70.
              pubmed: 12536217doi: 10.1038/nbt785google scholar: lookup

            Citations

            This article has been cited 60 times.
            1. Garcia-Rodriguez C, Yan S, Geren IN, Knopp KA, Dong J, Sun Z, Lou J, Conrad F, Wen WH, Farr-Jones S, Smith TJ, Brown JL, Skerry JC, Smith LA, Marks JD. A Four-Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotypes C and D.. Toxins (Basel) 2021 Sep 10;13(9).
              doi: 10.3390/toxins13090641pubmed: 34564645google scholar: lookup
            2. Zuverink M, Barbieri JT. Resolving the Molecular Steps in Clostridial Neurotoxin Light Chain Translocation.. J Exp Neurol 2020;1(4):123-134.
              doi: 10.33696/Neurol.1.020pubmed: 33615314google scholar: lookup
            3. Pandya P, Sayers RO, Ting JP, Morshedian S, Torres C, Cudal JS, Zhang K, Fitchett JR, Zhang Q, Zhang FF, Wang J, Durbin JD, Carrillo JJ, Espada A, Broughton H, Qian Y, Afshar S. Integration of phage and yeast display platforms: A reliable and cost effective approach for binning of peptides as displayed on-phage.. PLoS One 2020;15(6):e0233961.
              doi: 10.1371/journal.pone.0233961pubmed: 32479512google scholar: lookup
            4. Fan Y, Barash JR, Conrad F, Lou J, Tam C, Cheng LW, Arnon SS, Marks JD. The Novel Clostridial Neurotoxin Produced by Strain IBCA10-7060 Is Immunologically Equivalent to BoNT/HA.. Toxins (Basel) 2019 Dec 20;12(1).
              doi: 10.3390/toxins12010009pubmed: 31877649google scholar: lookup
            5. Rasetti-Escargueil C, Popoff MR. Antibodies and Vaccines against Botulinum Toxins: Available Measures and Novel Approaches.. Toxins (Basel) 2019 Sep 12;11(9).
              doi: 10.3390/toxins11090528pubmed: 31547338google scholar: lookup
            6. Sirois AR, Deny DA, Baierl SR, George KS, Moore SJ. Fn3 proteins engineered to recognize tumor biomarker mesothelin internalize upon binding.. PLoS One 2018;13(5):e0197029.
              doi: 10.1371/journal.pone.0197029pubmed: 29738555google scholar: lookup
            7. Garcia-Rodriguez C, Razai A, Geren IN, Lou J, Conrad F, Wen WH, Farr-Jones S, Smith TJ, Brown JL, Skerry JC, Smith LA, Marks JD. A Three Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotype E Subtypes.. Toxins (Basel) 2018 Mar 1;10(3).
              doi: 10.3390/toxins10030105pubmed: 29494481google scholar: lookup
            8. Zhou Y, Zou H, Yau C, Zhao L, Hall SC, Drummond DC, Farr-Jones S, Park JW, Benz CC, Marks JD. Discovery of internalizing antibodies to basal breast cancer cells.. Protein Eng Des Sel 2018 Jan 1;31(1):17-28.
              doi: 10.1093/protein/gzx063pubmed: 29301020google scholar: lookup
            9. Ti J, Li Z, Li X, Lu Y, Diao Y, Li F. Identification of one B-cell epitope from NS1 protein of duck Tembusu virus with monoclonal antibodies.. PLoS One 2017;12(7):e0181177.
              doi: 10.1371/journal.pone.0181177pubmed: 28746401google scholar: lookup
            10. Yao G, Lam KH, Perry K, Weisemann J, Rummel A, Jin R. Crystal Structure of the Receptor-Binding Domain of Botulinum Neurotoxin Type HA, Also Known as Type FA or H.. Toxins (Basel) 2017 Mar 8;9(3).
              doi: 10.3390/toxins9030093pubmed: 28282873google scholar: lookup
            11. Alves GG, Machado-de-Ávila RA, Chávez-Olórtegui CD, Silva ROS, Lobato FCF. Mapping of the continuous epitopes displayed on the Clostridium perfringens type D epsilon-toxin.. Braz J Microbiol 2017 Jul-Sep;48(3):570-575.
              doi: 10.1016/j.bjm.2016.10.023pubmed: 28223027google scholar: lookup
            12. Wang X, Stapleton JA, Klesmith JR, Hewlett EL, Whitehead TA, Maynard JA. Fine Epitope Mapping of Two Antibodies Neutralizing the Bordetella Adenylate Cyclase Toxin.. Biochemistry 2017 Mar 7;56(9):1324-1336.
              doi: 10.1021/acs.biochem.6b01163pubmed: 28177609google scholar: lookup
            13. Gaiotto T, Hufton SE. Cross-Neutralising Nanobodies Bind to a Conserved Pocket in the Hemagglutinin Stem Region Identified Using Yeast Display and Deep Mutational Scanning.. PLoS One 2016;11(10):e0164296.
              doi: 10.1371/journal.pone.0164296pubmed: 27741319google scholar: lookup
            14. Fan Y, Barash JR, Lou J, Conrad F, Marks JD, Arnon SS. Immunological Characterization and Neutralizing Ability of Monoclonal Antibodies Directed Against Botulinum Neurotoxin Type H.. J Infect Dis 2016 May 15;213(10):1606-14.
              doi: 10.1093/infdis/jiv770pubmed: 26936913google scholar: lookup
            15. Li X, Pushko P, Tretyakova I. Recombinant Hemagglutinin and Virus-Like Particle Vaccines for H7N9 Influenza Virus.. J Vaccines Vaccin 2015 Jun;6(3).
              doi: 10.4172/2157-7560.1000287pubmed: 26523241google scholar: lookup
            16. Rosenfeld L, Shirian J, Zur Y, Levaot N, Shifman JM, Papo N. Combinatorial and Computational Approaches to Identify Interactions of Macrophage Colony-stimulating Factor (M-CSF) and Its Receptor c-FMS.. J Biol Chem 2015 Oct 23;290(43):26180-93.
              doi: 10.1074/jbc.M115.671271pubmed: 26359491google scholar: lookup
            17. Fan Y, Dong J, Lou J, Wen W, Conrad F, Geren IN, Garcia-Rodriguez C, Smith TJ, Smith LA, Ho M, Pires-Alves M, Wilson BA, Marks JD. Monoclonal Antibodies that Inhibit the Proteolytic Activity of Botulinum Neurotoxin Serotype/B.. Toxins (Basel) 2015 Aug 26;7(9):3405-23.
              doi: 10.3390/toxins7093405pubmed: 26343720google scholar: lookup
            18. Fan Y, Geren IN, Dong J, Lou J, Wen W, Conrad F, Smith TJ, Smith LA, Ho M, Pires-Alves M, Wilson BA, Marks JD. Monoclonal Antibodies Targeting the Alpha-Exosite of Botulinum Neurotoxin Serotype/A Inhibit Catalytic Activity.. PLoS One 2015;10(8):e0135306.
              doi: 10.1371/journal.pone.0135306pubmed: 26275214google scholar: lookup
            19. Maslanka SE, Lúquez C, Dykes JK, Tepp WH, Pier CL, Pellett S, Raphael BH, Kalb SR, Barr JR, Rao A, Johnson EA. A Novel Botulinum Neurotoxin, Previously Reported as Serotype H, Has a Hybrid-Like Structure With Regions of Similarity to the Structures of Serotypes A and F and Is Neutralized With Serotype A Antitoxin.. J Infect Dis 2016 Feb 1;213(3):379-85.
              doi: 10.1093/infdis/jiv327pubmed: 26068781google scholar: lookup
            20. Kowalsky CA, Klesmith JR, Stapleton JA, Kelly V, Reichkitzer N, Whitehead TA. High-resolution sequence-function mapping of full-length proteins.. PLoS One 2015;10(3):e0118193.
              doi: 10.1371/journal.pone.0118193pubmed: 25790064google scholar: lookup
            21. Rojas G, Tundidor Y, Infante YC. High throughput functional epitope mapping: revisiting phage display platform to scan target antigen surface.. MAbs 2014;6(6):1368-76.
              doi: 10.4161/mabs.36144pubmed: 25484050google scholar: lookup
            22. Zhao L, Qu L, Zhou J, Sun Z, Zou H, Chen YY, Marks JD, Zhou Y. High throughput identification of monoclonal antibodies to membrane bound and secreted proteins using yeast and phage display.. PLoS One 2014;9(10):e111339.
              doi: 10.1371/journal.pone.0111339pubmed: 25353955google scholar: lookup
            23. Jenko KL, Zhang Y, Kostenko Y, Fan Y, Garcia-Rodriguez C, Lou J, Marks JD, Varnum SM. Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins.. Analyst 2014 Oct 21;139(20):5093-102.
              doi: 10.1039/c4an01270dpubmed: 25112421google scholar: lookup
            24. Abbott WM, Damschroder MM, Lowe DC. Current approaches to fine mapping of antigen-antibody interactions.. Immunology 2014 Aug;142(4):526-35.
              doi: 10.1111/imm.12284pubmed: 24635566google scholar: lookup
            25. Infante YC, Pupo A, Rojas G. A combinatorial mutagenesis approach for functional epitope mapping on phage-displayed target antigen: application to antibodies against epidermal growth factor.. MAbs 2014 May-Jun;6(3):637-48.
              doi: 10.4161/mabs.28395pubmed: 24589624google scholar: lookup
            26. Strotmeier J, Mahrhold S, Krez N, Janzen C, Lou J, Marks JD, Binz T, Rummel A. Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A.. FEBS Lett 2014 Apr 2;588(7):1087-93.
              doi: 10.1016/j.febslet.2014.02.034pubmed: 24583011google scholar: lookup
            27. Yin CH, Qin LT, Sun MY, Gao YL, Qi XL, Gao HL, Wang YQ, Wang XM. Antigenic analysis of monoclonal antibodies against different epitopes of σB protein of avian reovirus.. PLoS One 2013;8(11):e81533.
              doi: 10.1371/journal.pone.0081533pubmed: 24312314google scholar: lookup
            28. Sharon J, Rynkiewicz MJ, Lu Z, Yang CY. Discovery of protective B-cell epitopes for development of antimicrobial vaccines and antibody therapeutics.. Immunology 2014 May;142(1):1-23.
              doi: 10.1111/imm.12213pubmed: 24219801google scholar: lookup
            29. Mata-Fink J, Kriegsman B, Yu HX, Zhu H, Hanson MC, Irvine DJ, Wittrup KD. Rapid conformational epitope mapping of anti-gp120 antibodies with a designed mutant panel displayed on yeast.. J Mol Biol 2013 Jan 23;425(2):444-56.
              doi: 10.1016/j.jmb.2012.11.010pubmed: 23159556google scholar: lookup
            30. Gera N, Hill AB, White DP, Carbonell RG, Rao BM. Design of pH sensitive binding proteins from the hyperthermophilic Sso7d scaffold.. PLoS One 2012;7(11):e48928.
              doi: 10.1371/journal.pone.0048928pubmed: 23145025google scholar: lookup
            31. Chen C, Wang S, Wang H, Mao X, Zhang T, Ji G, Shi X, Xia T, Lu W, Zhang D, Dai J, Guo Y. Potent neutralization of botulinum neurotoxin/B by synergistic action of antibodies recognizing protein and ganglioside receptor binding domain.. PLoS One 2012;7(8):e43845.
              doi: 10.1371/journal.pone.0043845pubmed: 22952786google scholar: lookup
            32. Zhang Y, Lou J, Jenko KL, Marks JD, Varnum SM. Simultaneous and sensitive detection of six serotypes of botulinum neurotoxin using enzyme-linked immunosorbent assay-based protein antibody microarrays.. Anal Biochem 2012 Nov 15;430(2):185-92.
              doi: 10.1016/j.ab.2012.08.021pubmed: 22935296google scholar: lookup
            33. Meng Q, Garcia-Rodriguez C, Manzanarez G, Silberg MA, Conrad F, Bettencourt J, Pan X, Breece T, To R, Li M, Lee D, Thorner L, Tomic MT, Marks JD. Engineered domain-based assays to identify individual antibodies in oligoclonal combinations targeting the same protein.. Anal Biochem 2012 Nov 15;430(2):141-50.
              doi: 10.1016/j.ab.2012.08.005pubmed: 22922799google scholar: lookup
            34. Zdanovsky A, Zdanovsky D, Zdanovskaia M. Epitope mapping of botulinum neurotoxins light chains.. Toxicon 2012 Dec 1;60(7):1277-86.
              doi: 10.1016/j.toxicon.2012.08.002pubmed: 22922018google scholar: lookup
            35. Zhou Y, Marks JD. Discovery of internalizing antibodies to tumor antigens from phage libraries.. Methods Enzymol 2012;502:43-66.
              doi: 10.1016/B978-0-12-416039-2.00003-3pubmed: 22208981google scholar: lookup
            36. Makiya M, Dolan M, Agulto L, Purcell R, Chen Z. Structural basis of anthrax edema factor neutralization by a neutralizing antibody.. Biochem Biophys Res Commun 2012 Jan 6;417(1):324-9.
              doi: 10.1016/j.bbrc.2011.11.108pubmed: 22155239google scholar: lookup
            37. Corbett CR, Ballegeer E, Weedmark KA, Elias MD, Al-Saleem FH, Ancharski DM, Simpson LL, Berry JD. Epitope characterization of sero-specific monoclonal antibody to Clostridium botulinum neurotoxin type A.. Hybridoma (Larchmt) 2011 Dec;30(6):503-10.
              doi: 10.1089/hyb.2011.0032pubmed: 22149274google scholar: lookup
            38. Kalb SR, Santana WI, Geren IN, Garcia-Rodriguez C, Lou J, Smith TJ, Marks JD, Smith LA, Pirkle JL, Barr JR. Extraction and inhibition of enzymatic activity of botulinum neurotoxins /B1, /B2, /B3, /B4, and /B5 by a panel of monoclonal anti-BoNT/B antibodies.. BMC Biochem 2011 Nov 15;12:58.
              doi: 10.1186/1471-2091-12-58pubmed: 22085466google scholar: lookup
            39. Thanongsaksrikul J, Chaicumpa W. Botulinum neurotoxins and botulism: a novel therapeutic approach.. Toxins (Basel) 2011 May;3(5):469-88.
              doi: 10.3390/toxins3050469pubmed: 22069720google scholar: lookup
            40. Meng Q, Li M, Silberg MA, Conrad F, Bettencourt J, To R, Huang C, Ma J, Meyer K, Shimizu R, Cao L, Tomic MT, Marks JD. Domain-based assays of individual antibody concentrations in an oligoclonal combination targeting a single protein.. Anal Biochem 2012 Feb 15;421(2):351-61.
              doi: 10.1016/j.ab.2011.09.030pubmed: 22037290google scholar: lookup
            41. Sun EC, Ma JN, Liu NH, Yang T, Zhao J, Geng HW, Wang LF, Qin YL, Bu ZG, Yang YH, Lunt RA, Wang LF, Wu DL. Identification of two linear B-cell epitopes from West Nile virus NS1 by screening a phage-displayed random peptide library.. BMC Microbiol 2011 Jul 6;11:160.
              doi: 10.1186/1471-2180-11-160pubmed: 21729328google scholar: lookup
            42. Han T, Sui J, Bennett AS, Liddington RC, Donis RO, Zhu Q, Marasco WA. Fine epitope mapping of monoclonal antibodies against hemagglutinin of a highly pathogenic H5N1 influenza virus using yeast surface display.. Biochem Biophys Res Commun 2011 Jun 3;409(2):253-9.
              doi: 10.1016/j.bbrc.2011.04.139pubmed: 21569761google scholar: lookup
            43. Gray SA, Barr JR, Kalb SR, Marks JD, Baird CL, Cangelosi GA, Miller KD, Feldhaus MJ. Synergistic capture of Clostridium botulinum type A neurotoxin by scFv antibodies to novel epitopes.. Biotechnol Bioeng 2011 Oct;108(10):2456-67.
              doi: 10.1002/bit.23196pubmed: 21538339google scholar: lookup
            44. Xu W, Han L, Lin Z. Screening of random peptide library of hemagglutinin from pandemic 2009 A(H1N1) influenza virus reveals unexpected antigenically important regions.. PLoS One 2011 Mar 18;6(3):e18016.
              doi: 10.1371/journal.pone.0018016pubmed: 21437206google scholar: lookup
            45. Sun EC, Zhao J, Yang T, Liu NH, Geng HW, Qin YL, Wang LF, Bu ZG, Yang YH, Lunt RA, Wang LF, Wu DL. Identification of a conserved JEV serocomplex B-cell epitope by screening a phage-display peptide library with a mAb generated against West Nile virus capsid protein.. Virol J 2011 Mar 6;8:100.
              doi: 10.1186/1743-422X-8-100pubmed: 21375771google scholar: lookup
            46. Garcia-Rodriguez C, Geren IN, Lou J, Conrad F, Forsyth C, Wen W, Chakraborti S, Zao H, Manzanarez G, Smith TJ, Brown J, Tepp WH, Liu N, Wijesuriya S, Tomic MT, Johnson EA, Smith LA, Marks JD. Neutralizing human monoclonal antibodies binding multiple serotypes of botulinum neurotoxin.. Protein Eng Des Sel 2011 Mar;24(3):321-31.
              doi: 10.1093/protein/gzq111pubmed: 21149386google scholar: lookup
            47. Lollier V, Denery-Papini S, Larré C, Tessier D. A generic approach to evaluate how B-cell epitopes are surface-exposed on protein structures.. Mol Immunol 2011 Jan;48(4):577-85.
              doi: 10.1016/j.molimm.2010.10.011pubmed: 21111484google scholar: lookup
            48. Kalb SR, Garcia-Rodriguez C, Lou J, Baudys J, Smith TJ, Marks JD, Smith LA, Pirkle JL, Barr JR. Extraction of BoNT/A, /B, /E, and /F with a single, high affinity monoclonal antibody for detection of botulinum neurotoxin by Endopep-MS.. PLoS One 2010 Aug 17;5(8):e12237.
              doi: 10.1371/journal.pone.0012237pubmed: 20808925google scholar: lookup
            49. Scott N, Qazi O, Wright MJ, Fairweather NF, Deonarain MP. Characterisation of a panel of anti-tetanus toxin single-chain Fvs reveals cooperative binding.. Mol Immunol 2010 Jun;47(10):1931-41.
              doi: 10.1016/j.molimm.2010.02.020pubmed: 20413159google scholar: lookup
            50. Dong J, Thompson AA, Fan Y, Lou J, Conrad F, Ho M, Pires-Alves M, Wilson BA, Stevens RC, Marks JD. A single-domain llama antibody potently inhibits the enzymatic activity of botulinum neurotoxin by binding to the non-catalytic alpha-exosite binding region.. J Mol Biol 2010 Apr 9;397(4):1106-18.
              doi: 10.1016/j.jmb.2010.01.070pubmed: 20138889google scholar: lookup
            51. Cheng LW, Stanker LH, Henderson TD 2nd, Lou J, Marks JD. Antibody protection against botulinum neurotoxin intoxication in mice.. Infect Immun 2009 Oct;77(10):4305-13.
              doi: 10.1128/IAI.00405-09pubmed: 19651864google scholar: lookup
            52. Warner MG, Grate JW, Tyler A, Ozanich RM, Miller KD, Lou J, Marks JD, Bruckner-Lea CJ. Quantum dot immunoassays in renewable surface column and 96-well plate formats for the fluorescence detection of botulinum neurotoxin using high-affinity antibodies.. Biosens Bioelectron 2009 Sep 15;25(1):179-84.
              doi: 10.1016/j.bios.2009.06.031pubmed: 19643593google scholar: lookup
            53. Kalb SR, Lou J, Garcia-Rodriguez C, Geren IN, Smith TJ, Moura H, Marks JD, Smith LA, Pirkle JL, Barr JR. Extraction and inhibition of enzymatic activity of botulinum neurotoxins/A1, /A2, and /A3 by a panel of monoclonal anti-BoNT/A antibodies.. PLoS One 2009;4(4):e5355.
              doi: 10.1371/journal.pone.0005355pubmed: 19399171google scholar: lookup
            54. Scotcher MC, McGarvey JA, Johnson EA, Stanker LH. Epitope characterization and variable region sequence of f1-40, a high-affinity monoclonal antibody to botulinum neurotoxin type a (Hall strain).. PLoS One 2009;4(3):e4924.
              doi: 10.1371/journal.pone.0004924pubmed: 19290051google scholar: lookup
            55. Pai JC, Sutherland JN, Maynard JA. Progress towards recombinant anti-infective antibodies.. Recent Pat Antiinfect Drug Discov 2009 Jan;4(1):1-17.
              doi: 10.2174/157489109787236319pubmed: 19149692google scholar: lookup
            56. Nakouzi A, Rivera J, Rest RF, Casadevall A. Passive administration of monoclonal antibodies to anthrolysin O prolong survival in mice lethally infected with Bacillus anthracis.. BMC Microbiol 2008 Sep 23;8:159.
              doi: 10.1186/1471-2180-8-159pubmed: 18811967google scholar: lookup
            57. Adekar SP, Takahashi T, Jones RM, Al-Saleem FH, Ancharski DM, Root MJ, Kapadnis BP, Simpson LL, Dessain SK. Neutralization of botulinum neurotoxin by a human monoclonal antibody specific for the catalytic light chain.. PLoS One 2008 Aug 20;3(8):e3023.
              doi: 10.1371/journal.pone.0003023pubmed: 18714390google scholar: lookup
            58. Zarebski LM, Vaughan K, Sidney J, Peters B, Grey H, Janda KD, Casadevall A, Sette A. Analysis of epitope information related to Bacillus anthracis and Clostridium botulinum.. Expert Rev Vaccines 2008 Feb;7(1):55-74.
              doi: 10.1586/14760584.7.1.55pubmed: 18251694google scholar: lookup
            59. Baldwin MR, Tepp WH, Przedpelski A, Pier CL, Bradshaw M, Johnson EA, Barbieri JT. Subunit vaccine against the seven serotypes of botulism.. Infect Immun 2008 Mar;76(3):1314-8.
              doi: 10.1128/IAI.01025-07pubmed: 18070903google scholar: lookup
            60. Gai SA, Wittrup KD. Yeast surface display for protein engineering and characterization.. Curr Opin Struct Biol 2007 Aug;17(4):467-73.
              doi: 10.1016/j.sbi.2007.08.012pubmed: 17870469google scholar: lookup
            Subscribe to our newsletter
            Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.