Abstract
An epitope can be simply defined as that part of an antigen involved in its recogmtion by an antibody In the case of protein antigens, an epitope would consist of a group of mdlvidual ammo-acid side-chains close together on the protein surface Epitope mapping, then, becomes the process of locating the epitope, or identifying the mdividual ammo-acids mvolved Apart from its mtrmsic value for understanding protein structure-function relationships, it also has a practical value in generating antibody probes of defined specificity as research tools and in helping to define the immune response to pathogenic proteins and organisms Some authors have even extended the epitope concept to the mteraction between peptide hormones and then receptors (1), not every immunologist would be happy about this, but it does make the point that in mapping epitopes, we are studying a biological process of fundamental importance, that of protein-protein mteraction Epitope mapping is usually done with monoclonal antibodies (MAbs), though it can be done with polyclonal antisera in a rather less rigorous way, bearing in mind that antisera behave as mixtures of MAbs. Mapping can be done directly by X-ray crystallography of antibody-antigen complexes, but it can also be done by changing individual ammo-acids, by using antigen fragments and synthetic peptides or by competition methods in which two or more antibodies compete for the same, or adjacent, epitopes. The term “epitope mapping” has also been used to describe the attempt to determine all the major sites on a protein surface that can elicit an antibody response, at the end of which one might claim to have produced an “epitope map” of the protein mununogen (2) This mformatron might be very useful, for example, to someone wishing to produce antiviral vaccines. However, there is a limit to how far one can go down this road, because the map obtained may be influenced by how MAbs are selected and by the mapping method used Furthermore, the more strictly correct definmon of epltope mapping is based on antigenicity (the ability to recognize a specific antibody).
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Wells, J A (1995) Structural and functional epltopes in the growth hormone receptor complex Brotechnology 13,647–651
Atassl, M Z (1984) Antigenic structure of proteins Eur J Biochem 145, 1–20
Berzoksky, J A (1985) Intrinsic and extrmsic factors in protein antigenic structure Science 219,932–940
Reece, J C., Geysen, H M, and Rodda, S J. (1993) Mapping the major human T helper epltopes of tetanus toxm The emerging picture J Immunol 151,6175–6184
Clark, E A and Ledbetter, J A (1994) How B and T cells talk to each other Nature 367, 425–428
Howard, J C (1993) Restrlctlons on the use ofantlgemcpeptides by the nnmune system Proc Natl Acad Sci USA 90, 3777–3779
Stern, L J and Wiley, D C (1994) Antigenic peptide binding by class I and class II histocompatlbihty proteins Structure 2,245–251
Rammensee, H-G (1995) Chemistry of peptldes associated with MHC class I and class II molecules Curr Opin lmmunol 7, 85–96.
van Regenmortel, M H V (1989) Structural and functional approaches to the study of protein antlgemclty Immunol Today 10, 266–272
Barlow, D J, Edwards, M S, and Thornton, J M (1986) Continuous and discontinuous protein antigenic determinants Nature 322, 747–748
Laver, W G, An-, G M, Webster, R G, and Smith-Gill, S. J (1990) Epltopes on protein antigens misconceptions and reahties Cell 61, 553–556
Amit P, Mariuzza, R, Phllips, S, and Poljak, R. (1986) Three-dlmenslonal structure of an antigen-antibody complex Science 233, 747–753
Zvl, A, Kustanovlch, I, Felgelson, D, Levy, R, Elsenstem, M, Matsushita, S, Richalet-Secordel, P, van Regenmortel, M H V, and Anghster, J (1995) NMR mapping of the antlgenic determmant recogmsed by an anti-gp120, human immunodeficlency virus neutrahzing antibody Eur J Blochem 229, 178–187
Dore, I, Weiss, E, Altschuh, D, and van Regenmortel, M H V (1988) Visuahzatlon by electron microscopy of the location of tobacco mosaic virus epltopes reacting with monoclonal antibodies Virology 162, 279–289
Tzartos, S J, Rand, D E, Emarson, B L, and Lmdstrom, J M (1981) Mapping of surface structures of Electrophorus acetylchoinie receptor using monoclonal antibodies J Biol Chem 256,8635–8645
Le Thlet Thanh, Nguyen thl Man, Buu Mat, Phan Ngoc Tran, Nguyen thl Vinh Ha, and Morris, G. E (1991) Structural relatlonshlps between hepatltls B surface antigen in human plasma and dimers of recombmant vaccme a monoclonal antibody study Virus Res 21, 141–154
MohnarO, G A and Eby, W C (1984) One antigen may form two precipltm lmes and two spurs when tested with two monoclonal antibodies by gel diffusion assays Mol Immunol 21,181–184
Johne, B, Gadnell, M, and Hansen, K (1993) Epltope mappmg and bmdmg kinetlcs of monoclonal antibodies studied by real time blospeclfic interaction analysis using surface plasmon resonance J Immunol Methods 160, 191–198
Alexander, H., Alexander, S, Getzoff, E D, Tamer, J A, Geysen, H M, and Lemer, R A (1992) Altering the antlgemclty of proteins Proc Natl Acad Sci USA 89, 3352–3356
Ikeda, M, Hamano, K, and Shlbata, T (1992) Epltope mapping of anti-recA protem IgGs by region specified polymerase chain reaction mutagenesis J Blol Chem 267, 6291–4296
Wang, L F, Hertzog, P J, Galams, M, Overall, M L, Wame, G J, and Lmnane, A W (1994) Structure-functton analysis of human IFN-alpha-mapping of a conformattonal epttope by homologue scannmg J Immunol 152, 705–715
Pmg, L H and Lemon, S M (1992) Antigemc structure of human hepatttis-A virus defined by analysts of escape mutants selected agamst murme monoclonal antibodies J Virol 66, 2208–2216
Nguyen thi Man, Cartwrtght, A J, Osborne, M, and Morris, G E (1991) Structural changes in the C-terminal region of human brain creatme kmase studted with monoclonal antibodies Biochim Biophys Acta 1076, 245–251
Burnens, A, Demotz, S, Corradm, G, Binz, H, and Bosshard, H R (1987) Epitope mapping by differential chemical moditicatton of free and antibody-bound antigen Science 235,780–783
Jemmerson, R and Paterson, Y (1986) Mapping antigemc sites on a protem anttgen by the proteolysts of anttgen-antibody complexes Science 232, 1001–1004
Zhao, Y and Chatt, B. T (1995) Protein epitope mapping by mass spectrometry Anal Chem 66,3723–3726
Mazzoni, M R, Malmski, J A, and Hamm, H E (1991) Structural analysts of rod GTP-binding protem, Gt Limtted proteolytic dtgestton pattern of Gt with four proteases defines monoclonal anttbody epitope J Biol Chem 266,14,072–14,081
Ellgaard, L, Holtet, T L, Moestrup, S K, Etzerodt, M, and Thogersen, H C (1995) Nested sets of protem fragments and then use in epitope mapping-charactertzation of the epitope for the S4D5 monoclonal anttbody bmdmg to receptor-associated protem J Immunol Meth 180,53–61
Morris, G E (1989) Monoclonal antibody studies of creatme kmase The ART epttope evidence for an intermediate in protein foldmg Biochem J 257, 461–469
Morris, G E and Nguyen thi Man (1992) Changes at the N-termmus of human brain creatme kinase during a transition between inactive folding intermediate and active enzyme Biochim Bzophys Acta 1120,233–238
Stanley, K K (1988) Epttope mapping using pEX Meth Mol Biol 4,351–361
Nguyen thi Man and Morris, G E (1993) Use of epttope hbrartes to identify exon-specttic monoclonal anttbodies for charactertzatton of altered dystrophms in muscular dystrophy Amer J Hum Genet 52, 1057–1066
Wang, L F, Du Plessis, D H, White, J R, Hyatt, A R, and Eaton, B T (1995) Use of a gene-targeted phage display random epttope library to map an antrgemc determinant on the bluetongue vnus outer capsid protein VP5 J Immunol Methods 178, 1–12
Lenstra, J A, Kusters, J G,and vander Zeijst, B A M (1990) Mapping of viral epttopes with procaryotic expression systems (review) Arch Virol 110,1–24
Thanh, L T, Nguyen tht Man, Hart, S, Sewry, C A, Dubowitz V, and Moms, G E (1995) Characterization of genettc delettons in Becker Muscular Dystrophy using monoclonal antibodies against a deletion-prone region of dystrophm Amer J Med Genet 58, 177–186
Burch, H B, Nagy, E V, Kam, K C, Lanar, D E, Carr, F E, Wartofsky, L, and Burman, K D(1993) Expression polymerase chain reaction for the in vttro synthesis and epttope mapping of autoanttgen Appbcatton to the human thyrotropm receptor f Immunol Methods 158,123–130
Sedgwtck, S G, Nguyen tht Man, Ellis, J M., Crowne, H, and Moms, G E (1991) Rapid mapping by transposon mutagenesis of epttopes on the muscular dystrophy protein, dystrophm Nucietc Acids Res 19,5889–5894
Frtguet, B, Fedorov, A N, and DJavadi-Ohamance, L (1993) In vitro gene expression for the localtzatton of antigemc determmants-apphcation to the E Coli tryptophan synthase beta2 subumt. J Immunol Methods 158, 243–249
Gross, C H and Rohrmann, G F (1990) Mapping unprocessed epitopes using deletion mutagenesis of gene fusions Biotechniques 8, 196–202
Geysen, H M, Meleon, R H, and Barteling, S J (1984) Use of peptlde synthesis to probe viral antigens for epttopes to a resolution of a single amino-acid Proc Nutl Acad Sci USA 81, 3998–4002
Frank, R, Kless, M, Lahmann, H, Behn, C H, and Gausepohl, H (1995) Combmatorlal synthesis on membrane supports by the SPOT techmque, in Peptides 1994 (Mala, H L S, ed), ESCOM, Leaden, pp 479–480
Holmes, C P, Adams, C L, Kochersperger, L M, Mortensen, R B, and Aldwin, LA (1995) The use of hght-directed combmatonal peptlde synthesis in epltope mapping Biopolymers 37, 199–211
Houghten, R A, Pmllla, C, Blondelle, S E, Appel, J R, Dooley, C T, and Cuervo, J H (1991) Generatlon and use of synthetic peptlde combmatorlal libraries for basic research and drug discovery Nature 354, 84–86
Scott, J K and Smith, G P (1990) Searching for peptlde hgands with an epltope library Science 249, 386–390
Morris, G E, Nguyen, C, and Nguyen thl Man (1995) Speclficlty and V, sequence of two monoclonal antibodies against the N-termmus of dystrophin Biochem J 78,355–359
Mattheakls, J C, Bhatt, R R, and Dower, W J (1994) An in vitro display system for ldentlfymg hgands from very large peptlde hbrarles Proc Natl Acad Sci USA 91, 9022–9026
Lu, Z., Murray, K S, van Cleave, V, LaVallle, E R, Stahl, M L., and McCoy, J M (1995) Expression of thloredoxin random peptlde hbrarles on the Escherlchla co11 cell surface as functional fusions to flagellin Bio/technology 13, 366–372
Moms, G E (ed.) (1996), Methods in Molecular Bzology, vol 66 Epztope Mapping Protocols Humana, Totowa, NJ
Wang, K S and Strauss, J H (1991) Use of a lambda gtl 1 expression library to locahze a neutrahzing antibody-bmdmg site in glycoprotem-E2 of Smdbls virus J Virol 65,7037–7040
Ho, D D, Fung, M S C, Cao, Y Z, LI, X L., Sun, C, Chang, T W, and Sun, N C (1991) Another dlscontmuous epltope on glycoprotem gp120 that IS important in Human Immunodeficlency Virus Type-l neutrahzatlon is identified by a monoclonal antlbody Proc Natl Acad Sci USA 88, 8949–8952
Rowlands, D J (1992) How can peptlde vaccines work FEMS Microbiol Lett 100, 479–481
Albam, S and Roudler, J (1992) Molecular basis for the assoclatlon between hladr4 and rheumatoid arthritis-from the shared epltope hypothesis to a peptldlc model of rheumatold arihrltls Clin Biochem 25,209–212
Butler, M H, Sohmena, M, Dirkx, R, Hayday, A, and Decamllh, P (I993) Identificatlon of a dominant epltope of glutamic acid decarboxylase (GAD-65) recognized by autoantlbodles in Stiff-Man syndrome J Exp Med 178,2097–2106
Carson, D A (1994) The value of epltope mappmg in autoimmune diseases J Clin Invest 94, 1713
Frank, M B, Itoh, K, and McCubbm, V (1994) Epltope mappmg of the 52-kD Ro/SSA autoantigen Clin Exp Immunol 95,390–396
Palace, J, Vincent, A, Beeson, D, and Newsom-Davis, J (1994) Immunogenicity of human recombinant acetylcholme receptor alpha subumt cytoplasmlc epltopes dommate the antibody response in four mouse strams Autoimmunity 18, 113–119
Lm, M S, Ma, Y H, Hayden, M R, and Brunzell, J D (1992) Mapping of the epltope on lrpoprotem llpase recogmzed by a monoclonal antibody (5D2) which mhlblts hpase actlvlty Biochim Biophys Acta 1128, 113–115
Ptetu, G, Rtbba, A S, Cherel, G, and Meyer, D (1992) Epttope mappmg by cDNA expression of a monoclonal antibody which mhtbits the bmdmg of von Wtllebrand factor to platelet glycoprotem-IIblIIIa Biochem J 284,711–715
Landis, R C, Bennett, R I, and Hogg, N (1993) A novel LFA-1 acttvation epttope maps to the I-domain J Cell Btol 120, 1519–1527
Mans, C A, Underwood, P A, Bean, P A, Sheehan, M, and Charlesworth, J A (1994) Relative topography of btologtcally active domains of human vttronectm-evidence from monoclonal antibody epttope and denaturatton studtes J Blol Chem 269, 23,845–23,852
Melhus, H, Bavik, C O, Rask, L, Peterson, P A, and Ertksson, U (1995) Epttope mapping of a monoclonal antibody that blocks the bmdmg of retmol-binding protem to its receptor Brochem Bzophys Res Commun 210, 105–112
Morns, G E, Frost, L C, Newport, P A, and Hudson, N (1987) Monoclonal antibody studies of creatme kmase Antibody-binding sites in the N-terminal regton of creatme kmase and effects of anttbody on enzyme refolding Blochem J 248, 53–59
Syu, W J and Kahan, L (1992) Both ends of Eschertchra Cob rtbosomal protein-S 13 are tmmunochemically accesstble in sttu J Protein Chem 11,225–230
Nmg, G, Maunsbach, A B, Lee, Y J, and Moller, J V (1993) Topology of Na,K-ATPase alpha subumt epltopes analyzed with oltgopeptide-specific antibodies and double-labelmg nnmunoelectron mtcroscopy FEBS Lett 336,521–524
Morris, G E and Cartwright, A J (1990) Monoclonal antibody studies suggest a catalytic site at the interface between domams in creatme kmase Blochim Bzophys Acta 1039,318–322
Kruger, M, Wright, J, and Wang, K (1991) Nebulm as a length regulator of thm filaments of vertebrate skeletal muscles-correlatton of thm filament length, nebulm size, and epitopeprofile J Cell Biol 115,97–107
Morris, G E, Snnmons, C, and Nguyen tht Man (1995) Apo-dystrophms (Dp 140 and Dp71) and dystrophm sphcmg isoforms in developmg brain. Biochem Btophys Res Commun 215, 361–367
Carter, J M (1994) Epttope predictton methods, in Peptrde Analyszs Protocols (Dunn, B M and Pennmgton, M W, eds), Humana, Totowa, NJ, pp 193–206
Mandal, C, Kmgery, B D, Anchm, J M, Subramantam, S, and Lmthtcum, D S (1996) ABGEN. a knowledge-based automated approach for anttbody structure modellmg Nature Blotechnol 14,323–328.
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Morris, G.E. (1998). Epitope Mapping. In: Rapley, R., Walker, J.M. (eds) Molecular Biomethods Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-59259-642-3_47
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