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Improving the affinity of antigens for mutated antibodies by use of statistical molecular design
Author(s) -
Mandrika Ilona,
Prusis Peteris,
Bergström Jakob,
Yahorava Sviatlana,
Wikberg Jarl E. S.
Publication year - 2008
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.999
Subject(s) - peptide , antigen , epitope , antibody , chemistry , peptide library , complementarity determining region , microbiology and biotechnology , computational biology , biology , biochemistry , peptide sequence , monoclonal antibody , gene , genetics
We demonstrate the use of statistical molecular design (SMD) in the selection of peptide libraries aimed to systematically investigate antigen‐antibody binding spaces. Earlier, we derived two novel antibodies by mutating the complementarity‐determining region of the anti‐p24 (HIV‐1) single chain Fv antibody, CB4‐1 that had lost their affinity for a p24 epitope‐homologous peptide by 8‐ and 60‐fold. The present study was devoted to explore how peptide libraries can be designed under experimental design criteria for effective screening of peptide antigens. Several small peptide–antigen libraries were selected using SMD principles and their activities were evaluated by their binding to SPOT‐synthesized peptide membranes and by fluorescence polarization (FP). The approach was able to reveal the most critical residues required for antigen binding, and finally to increase the binding activity by proper modifications of amino acids in the peptide antigen. A model of the active peptide binding pocket formed by the mutated scFv and the antigen was compatible with the information gained from the experimental data. Our results suggest that SMD approaches can be used to explore peptide antigen features essential for their interactions with antibodies. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd.