Affinity maturation by semi-rational approaches.
Rational engineering methods can be applied with success to optimize physicochemical characteristics of antibodies. Application of in silico analysis and prediction methods to antibody Fv regions can help to find residues affecting antibody-antigen affinity when high-resolution antibody structures or antibody-antigen complex structures are known. In these cases, the identification of residues affecting affinity can facilitate the selection of candidates for guided maturation by PCR using degenerate oligonucleotides. Here, we describe the utilization of a semi-rational approach to enhance the affinity of antibodies by combining in silico and traditional wet lab-based methods.
Centro de Investigaviones Biológicas, Madrid, Spain.
Rodrigo
R
Barderas
Johan
J
Desmet
Philippe
P
Alard
J Ignacio
JI
Casal
D000596:Amino Acids / Q000235:genetics; D000915:Antibody Affinity / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000276:immunology; D000941:Antigens / Q000276:immunology; D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005755:Gastrins / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology; D008958:Models, Molecular; D008967:Molecular Biology / Q000379:methods*; D009154:Mutation / Q000235:genetics; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D057127:Single-Chain Antibodies / Q000276:immunology
D000596:Amino Acids; D000936:Antigen-Antibody Complex; D000941:Antigens; D000939:Epitopes; D005755:Gastrins; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D057127:Single-Chain Antibodies; C013550:gastrin 17