Abstract
If antibody-based therapeutics are to be rationally designed to give optimal interactions with their target antigens, it is essential to understand these antibody/antigen interactions as fully as possible. This has been greatly advanced by the recent developments in mass spectrometry (MS), structural identification, and bioinformatics, which allow the relatively simple and rapid characterization of protein—protein interactions in a high-throughput manner. Previous methods of mapping an antibody-binding site (epitope) on an antigen have relied on peptide libraries (1). Such methods are suitable for the identification of continuous amino acid sequences (2) but not for those that are conformationally dependent. This latter group forms a major category of antibody/antigen interactions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Benjamin, D. C., Berzofsky, J. A., East, I. J., et al. (1984) The antigenic structure of proteins: a reappraisal. Annu. Rev. Immunol. 2, 67–101.
Berzofsky, J. A. (1985) Intrinsic and extrinsic factors in protein antigenic structure. Science 229, 932–940.
Jemmerson, R. and Paterson, Y. (1986) Mapping epitopes on a protein antigen by the proteolysis of antigen-antibody complexes. Science 232, 1001–1004.
Eisenberg, R. J., Long, D., Pereira, L., Hampar, B., Zweig, M., and Cohen, G. H. (1982) Effect of monoclonal antibodies on limited proteolysis of native glycoprotein gD of herpes simplex virus type 1. J. Virol. 41, 478–488.
Moelling, K., Scott, A., Dittmar, K. E., and Owada, M. (1980) Effect of p15-associated protease from an avian RNA tumor virus on avian virus-specific polyprotein precursors. J. Virol. 33, 680–688.
Schwyzer, M., Weil, R., Frank, G., and Zuber, H. (1980) Amino acid sequence analysis of fragments generated by partial proteolysis from large simian virus 40 tumor antigen. J. Biol. Chem. 255, 5627–5634.
Sheshberadaran, H. and Payne, L. G. (1988) Protein antigen-monoclonal antibody contact sites investigated by limited proteolysis of monoclonal antibody-bound antigen: protein “footprinting.” Proc. Natl. Acad. Sci. USA 85, 1–5.
Suckau, D., Kohl, J., Karwath, G., et al. (1990) Molecular epitope identification by limited proteolysis of an immobilized antigen-antibody complex and mass spectrometric peptide mapping. Proc. Natl. Acad. Sci. USA 87, 9848–9852.
Van de, W. J., Deininger, S. O., Macht, M., Przybylski, M., and Gershwin, M. E. (1997) Detection of molecular determinants and epitope mapping using MALDI-TOF mass spectrometry. Clin. Immunol. Immunopathol. 85, 229–235.
Yi, J. and Skalka, A. M. (2000) Mapping epitopes of monoclonal antibodies against HIV-1 integrase with limited proteolysis and matrix-assisted laser desorption ionization time-offlight mass spectrometry. Biopolymers 55, 308–318.
Spencer, D. I., Robson, L., Purdy, D., et al. (2002) A strategy for mapping and neutralizing conformational immunogenic sites on protein therapeutics. Proteomies 2, 271–279.
Reid, G., Gan, B. S., She, Y. M., Ens, W., Weinberger, S., and Howard, J. C. (2002) Rapid identification of probiotic Lactobacillus biosurfactant proteins by ProteinChip tandem mass spectrometry tryptic peptide sequencing. Appl. Environ. Microbiol. 68, 977–980.
Huhalov, A. Spencer, D. I. R., Hawkins, R. E., Perkins, S. J., Begent, R. H. J., and Chester, K. A. (2002) Capture and analysis of interacting tagged-proteins: an application to map the carcinoembryonic antigen binding-site of single chain Fv molecule MFE-23. J. Mol. Biol.,in press.
Chester, K. A., Begent, R. H., Robson, L., et al. (1994) Phage libraries for generation of clinically useful antibodies. Lancet 343, 455–456.
Begent, R. H., Verhaar, M. J., Chester, K. A., et al. (1996) Clinical evidence of efficient tumor targeting based on single-chain Fv antibody selected from a combinatorial library. Nat. Med. 2, 979–984.
Mayer, A., Tsiompanou, E., O’Malley, D., et al. (2000) Radioimmunoguided surgery in colorectal cancer using a genetically engineered anti-CEA single-chain Fv antibody. Clin. Cancer Res. 6, 1711–1719.
Oikawa, S., Imajo, S., Noguchi, T., Kosaki, G., and Nakazato, H. (1987) The carcinoembryonic antigen (CEA) contains multiple immunoglobulin-like domains. Biochem. Biophvs. Res. Commun. 144, 634–642.
Paxton, R. J., Mooser, G., Pande, H., Lee, T. D., and Shively, J. E. (1987) Sequence analysis of carcinoembryonic antigen: identification of glycosylation sites and homology with the immunoglobulin supergene family. Proc. Natl. Acad. Sci. USA 84, 920–924.
Boehm, M. K., Mayans, M. O., Thornton, J. D., Begent, R. H., Keep, P. A., and Perkins, S. J. (1996) Extended glycoprotein structure of the seven domains in human carcinoembryonic antigen by X-ray and neutron solution scattering and an automated curve fitting procedure: implications for cellular adhesion. J. Mol. Biol. 259, 718–736.
Boehm, M. K., Corper, A. L., Wan, T., et al. (2000) Crystal structure of the anti(carcinoembryonic antigen) single-chain Fv antibody MFE-23 and a model for antigen binding based on intermolecular contacts. Biochem. J. 346, 519–528.
Boehm, M. K. and Perkins, S. J. (2000) Structural models for carcinoembryonic antigen and its complex with the single-chain Fv antibody molecule MFE23. FEES Lett. 475, 11–16.
Michael, N. P., Chester, K. A., Melton, R. G., et al. (1996) In vitro and in vivo characterisation of a recombinant carboxypeptidase G2::anti-CEA scFv fusion protein. Immunotechnology 2, 47–57.
Bagshawe, K. D. (1987) Antibody directed enzymes revive anti-cancer prodrugs concept. Br. J. Cancer 56, 531–532.
Rowsell, S., Pauptit, R. A., Tucker, A. D., Melton, R. G., Blow, D. M., and Brick, P. (1997) Crystal structure of carboxypeptidase G2, a bacterial enzyme with applications in cancer therapy. Structure 5, 337–347.
Hoogenboom, H. R. and Chames, P. (2000) Natural and designer binding sites made by phage display technology. Immunol. Today 21, 371–378.
Verma, R., Boleti, E., and George, A. J. T. (2000) Antibody engineering: comparison of bacterial, yeast, insect and mammalian expression systems. J. Immunol. Methods 216, 165–181.
Clauser, K. R., Baker, P., and Burlingame, A. L. (1999) Role of accurate mass measurement (+/– 10 ppm) in protein identification strategies employing MS or MS/MS and database searching. Anal. Chem. 71, 2871–2882.
Sayle, R. A. and Milner-White, E. J. (1995) RASMOL: biomolecular graphics for all. Trends Biochem. Sci. 20, 374.
Berman, H. M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T. N., Weissig, H., et al. (2000) The Protein Data Bank. Nucleic Acids Res. 28, 235–242.
Kabsch, W. and Sander, C. (1983) Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features. Biopolymers 22, 2577–2637.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Springer Science+Business Media New York
About this chapter
Cite this chapter
Huhalov, A., Spencer, D.I.R., Chester, K.A. (2003). A Strategy for Characterizing Antibody/Antigen Interactions Using ProteinChip® Arrays. In: Conn, P.M. (eds) Handbook of Proteomic Methods. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59259-414-6_9
Download citation
DOI: https://doi.org/10.1007/978-1-59259-414-6_9
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61737-504-0
Online ISBN: 978-1-59259-414-6
eBook Packages: Springer Book Archive