title: In vitro evolution of a neutralizing human antibody to human immunodeficiency virus type 1 to enhance affinity and broaden strain cross-reactivity.
abstract: A method is described that allows for the improvement of antibody affinity. This method, termed complementary-determining region (CDR) walking, does not require structural information on either antibody or antigen. Complementary-determining regions are targeted for random mutagenesis followed by selection for fitness, in this case increased binding affinity, by the phage-display approach. The current study targets a human CD4-binding-site anti-gp120 antibody that is potently and broadly neutralizing. Evolution of affinity of this antibody demonstrates in this case that affinity can be increased while reactivity to variants of human immunodeficiency virus type 1 is broadened. The neutralizing ability of this antibody is improved, as assayed with laboratory and primary clinical isolates of human immunodeficiency virus type 1. The ability to produce human antibodies of exceptional affinity and broad neutralizing ability has implications for the therapeutic and prophylactic application of antibodies for human immunodeficiency virus type 1 infection.
authors.affiliation: Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037.
authors.forename: C F
authors.initials: CF
authors.lastname: Barbas
authors.forename: D
authors.initials: D
authors.lastname: Hu
authors.forename: N
authors.initials: N
authors.lastname: Dunlop
authors.forename: L
authors.initials: L
authors.lastname: Sawyer
authors.forename: D
authors.initials: D
authors.lastname: Cababa
authors.forename: R M
authors.initials: RM
authors.lastname: Hendry
authors.forename: P L
authors.initials: PL
authors.lastname: Nara
authors.forename: D R
authors.initials: DR
authors.lastname: Burton
mesh_terms: D000595:Amino Acid Sequence; D000915:Antibody Affinity; D001483:Base Sequence; D001665:Binding Sites; D001666:Binding Sites, Antibody; D003429:Cross Reactions; D015483:HIV Antibodies / Q000235:genetics / Q000276:immunology*; D015488:HIV Antigens / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009500:Neutralization Tests; D013329:Structure-Activity Relationship
chemical_list: D015483:HIV Antibodies; D015488:HIV Antigens; D015699:HIV Envelope Protein gp120