title: Generation of a family-specific phage library of llama single chain antibody fragments that neutralize HIV-1.
abstract: Recently, we described llama antibody fragments (VHH) that can neutralize human immunodeficiency virus, type 1 (HIV-1). These VHH were obtained after selective elution of phages carrying an immune library raised against gp120 of HIV-1 subtype B/C CN54 with soluble CD4. We describe here a new, family-specific approach to obtain the largest possible diversity of related VHH that compete with soluble CD4 for binding to the HIV-1 envelope glycoprotein. The creation of this family-specific library of homologous VHH has enabled us to isolate phages carrying similar nucleotide sequences as the parental VHH. These VHH displayed varying binding affinities and neutralization phenotypes to a panel of different strains and subtypes of HIV-1. Sequence analysis of the homologs showed that the C-terminal three amino acids of the CDR3 loop were crucial in determining the specificity of these VHH for different subtype C HIV-1 strains. There was a positive correlation between affinity of VHH binding to gp120 of HIV-1 IIIB and the breadth of neutralization of diverse HIV-1 envelopes. The family-specific approach has therefore allowed us to better understand the interaction of the CD4-binding site antibodies with virus strain specificity and has potential use for the bioengineering of antibodies and HIV-1 vaccine development.
authors.affiliation: Division of Infection and Immunity, University College London, Medical Research Council/University College London Centre for Medical Molecular Virology, London W1T 4JF, United Kingdom.
authors.forename: Willie W L
authors.initials: WW
authors.lastname: Koh
authors.forename: Soren
authors.initials: S
authors.lastname: Steffensen
authors.forename: Maria
authors.initials: M
authors.lastname: Gonzalez-Pajuelo
authors.forename: Bart
authors.initials: B
authors.lastname: Hoorelbeke
authors.forename: Andrea
authors.initials: A
authors.lastname: Gorlani
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Szynol
authors.forename: Anna
authors.initials: A
authors.lastname: Forsman
authors.forename: Marlén M I
authors.initials: MM
authors.lastname: Aasa-Chapman
authors.forename: Hans
authors.initials: H
authors.lastname: de Haard
authors.forename: Theo
authors.initials: T
authors.lastname: Verrips
authors.forename: Robin A
authors.initials: RA
authors.lastname: Weiss
mesh_terms: D016915:AIDS Vaccines / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry; D001483:Base Sequence; D001665:Binding Sites; D015704:CD4 Antigens / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000737:chemistry; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D019151:Peptide Library; D012689:Sequence Homology, Nucleic Acid; D057127:Single-Chain Antibodies / Q000737:chemistry*
chemical_list: D016915:AIDS Vaccines; D000906:Antibodies; D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; D019151:Peptide Library; D057127:Single-Chain Antibodies
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