The relation between domain structures of recombinant monoclonal antibody fragments and their reaction kinetics was studied for the first time using a novel biosensor based on surface plasmon resonance technology. The association and dissociation rate constants of Fab, Fv and single domain (VH fragment) anti–lysozyme antibodies were determined and compared to the intact monoclonal antibody. Fab and Fv fragments showed similar reaction kinetics and had affinity constants of 6 × 109 M−1 and 25 × 109 M−1, respectively. The single domain antibody had significantly different reaction kinetics compared to the fragments consisting of paired heavy and light chain domains. The VH domain had both a higher dissociation and a lower association rate constant, which resulted in an affinity constant approximately 250 times lower than the Fab fragment. This rapid evaluation of antibody reaction kinetics should prove to be an important selection parameter when comparing antibody fragments for their utility in therapeutic or other applications.
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Foote, J. and Milstein, C. 1991. Kinetic maturation of an immune response. Nature 352: 530–532.
Winter, G. and Milstein, C. 1991. Man made antibodies. Nature 349: 293–299.
Ward, E.S., Güssow, D., Griffith, A.D., Jones, P.T. and Winter, G. 1989. Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341: 544–546.
Malmborg, A.-C., Michaëlsson, A., Ohlin, M., Jansson, B. and Borrebaeck, C.A.K. 1992. Real time analysis of antibody-antigen reaction kinetics. Scand. J. Immunol. In press.
Karlsson, R., Michaëlsson, A. and Mattsson, L. 1991. Kinetic analysis of monoclonal antibody-antigen interactions with a new biosensor based analytical system. J. Immunol. Methods 145: 229–240.
Fischmann, T.O., Bentley, G.A., Bhat, T.N., Boulot, G., Mariuzza, R.A., Phillips, S.E.V., Tello, D. and Poljak, R.J. 1991. Crystallo-graphic refinement of the three-dimensional structure of the FabD 1.3-lysozyme complex at 2.5-Å resolution. J. Biol. Chem. 266: 12915–12920.
Nygren, H., Czerkinsky, C. and Stenberg, M. 1985. Dissociation of antibodies to surface-immobilized antigen. J. Immunol. Methods 85: 87–95.
Nygren, H., Werthen, M. and Stenberg, M. 1987. Kinetics of antibody binding to solid-phase-immobilized antigen. Effect of diffusion rate limitation and steric interaction. J. Immunol. Methods 101: 63–71.
Mariuzza, R.A., Jankovic, D.L.J., Boulot, G., Amit, A.G., Saludjian, P., Le Guern, A., Mazie, J.C. and Poljak, R.J. 1983. Preliminary crystallographic study of the complex between the Fab fragment of a monoclonal anti-lysozyme antibody and its antigen. J. Mol. Biol. 170: 1055–1058.
Ward, E.S. 1991. Expression and purification of antibody fragments using Escherichia coli as a host, p. 121–137. In: Antibody Engineering. A Practical Guide. Borrebaeck, C. A. K. (Ed.). W. H. Freeman and Company, New York.
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Borrebaeck, C., Malmborg, AC., Furebring, C. et al. Kinetic Analysis of Recombinant Antibody–Antigen Interactions: Relation Between Structural Domains and Antigen Binding. Nat Biotechnol 10, 697–698 (1992). https://doi.org/10.1038/nbt0692-697