title: Selection of human anti-human immunodeficiency virus type 1 envelope single-chain antibodies from a peripheral blood cell-based phage repertoire.
abstract: Monoclonal antibodies play an important role in the development of diagnostic assays. Instead of using hybridoma technology to isolate human immunodeficiency virus type 1-specific antibodies, a phage-displayed antibody library was generated from a small number (10(7)) of peripheral blood lymphocytes from a seropositive donor. Two families of single-chain antibodies (scFvs) were selected by biopanning with the envelope precursor gp160. ELISA and competition in the BIAcore system revealed that one antibody family recognized a conformation-sensitive epitope within gp120, while the other antibody family was gp41-specific. The latter group had sequence similarity to antibodies recognizing the cluster III epitope of gp41. Binding of scFvs to gp160 could be inhibited with the donor's serum antibodies, indicating that antibodies with a similar specificity were circulating in the donor's blood. Competition experiments suggested that the epitope of the anti-gp41 antibodies was recognized by a broad range of patients' sera: 21 out of 22 sera from North American and all 20 sera from African seropositive patients inhibited binding of scFvs. In contrast, three sera from this panel did not react with the epitope of the anti-gp120 antibodies. These data indicate that, because of the conserved nature of its epitope, the anti-gp41 antibody will be suitable for diagnostic applications.
authors.affiliation: Biosciences Research Unit, Organon Teknika, Boxtel, The Netherlands.
authors.forename: J J
authors.initials: JJ
authors.lastname: de Haard
authors.forename: B
authors.initials: B
authors.lastname: Kazemier
authors.forename: P
authors.initials: P
authors.lastname: Oudshoorn
authors.forename: P
authors.initials: P
authors.lastname: Boender
authors.forename: B
authors.initials: B
authors.lastname: van Gemen
authors.forename: M J
authors.initials: MJ
authors.lastname: Koolen
authors.forename: G
authors.initials: G
authors.lastname: van der Groen
authors.forename: H R
authors.initials: HR
authors.lastname: Hoogenboom
authors.forename: J W
authors.initials: JW
authors.lastname: Arends
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages; D001667:Binding, Competitive; D015483:HIV Antibodies / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D019302:HIV Envelope Protein gp160 / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000276:immunology; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D017421:Sequence Analysis
chemical_list: D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; C055966:immunoglobulin Fv