title: Improving antibody binding affinity and specificity for therapeutic development.
abstract: Affinity maturation is an important part of the therapeutic antibody development process as in vivo activity often requires high binding affinity. Here, we describe a targeted approach for affinity improvement of therapeutic antibodies. Sets of CDR residues that are solvent accessible and relatively diverse in natural antibodies are targeted for diversification. Degenerate oligonucleotides are used to generate combinatorial phage-displayed antibody libraries with varying degree of diversity at randomized positions from which high-affinity antibodies can be selected. An advantage of using antibodies for therapy is their exquisite target specificity, which enables selective antigen binding and reduces off-target effects. However, it can be useful, and often it is necessary, to generate cross-reactive antibodies binding to not only the human antigen but also the corresponding non-human primate or rodent orthologs. Such cross-reactive antibodies can be used to validate the therapeutic targeting and examine the safety profile in preclinical animal models before committing to a costly development track. We show how affinity improvement and cross-species binding can be achieved in a one-step process.
authors.affiliation: Genentech Inc., San Francisco, CA, USA.
authors.forename: Jenny
authors.initials: J
authors.lastname: Bostrom
authors.forename: Chingwei V
authors.initials: CV
authors.lastname: Lee
authors.forename: Lauric
authors.initials: L
authors.lastname: Haber
authors.forename: Germaine
authors.initials: G
authors.lastname: Fuh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use*; D000915:Antibody Affinity / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D001483:Base Sequence; D002999:Clone Cells; D003062:Codon / Q000235:genetics; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D020128:Inhibitory Concentration 50; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D019151:Peptide Library
chemical_list: D000906:Antibodies; D003062:Codon; D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D009841:Oligonucleotides; D019151:Peptide Library