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Development of functional human monoclonal single-chain variable fragment antibody against hiv-1 from human cervical b cells

Academic Article
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Overview

authors

  • Berry, J. D.
  • Rutherford, J.
  • Silverman, G. J.
  • Kaul, R.
  • Elia, M.
  • Gobuty, S.
  • Fuller, R.
  • Plummer, F. A.
  • Barbas III, Carlos

publication date

  • April 2003

journal

  • Hybridoma and Hybridomics  Journal

abstract

  • A panel of novel recombinant single-chain variable fragment (scFv) antibody against human immunodeficiency virus type-1 (HIV-1) was isolated and characterized. We generated human scFvs using RNA harvested from cervical B lymphocytes of Kenyan prostitutes who are highly exposed to HIV-1, but remain persistently seronegative. The variable regions of the heavy (VH) and light (VL) chain antibody genes were selected as hybrids using guided-selection with the VL and VH, respectively, of a derivative of IgGb(12) using the phagemid vector pComb3X. IgGb(12) is a previously well-characterized HIV-1 neutralizing human monoclonal antibody (MAb). One of the hybrid scFv, IgA6/4L, neutralizes HIV-1 infectivity in in vitro cell culture assay. The cervical VH and VL chain antibody genes were connected by a DNA linker and subcloned in pComb3X. The cervical scFv clones were functional in recognizing HIV-1 gp120 by enzyme-linked immunosorbant assay (ELISA) and on cells in flow cytometry. Whole IgGb(12) does not inhibit binding of clones IgA6/5k nor IgA6/30lambda to gp120, which suggests that they bind different epitopes. Nucleotide sequence analysis of the cervical scFv show the clones are unique and reveal interesting characteristics of human cervical V gene pools. This work demonstrates, for the first time, cloning of a functional scFv MAb to a sexually transmitted disease pathogen from local cervical B-cell pools in exposed humans.

subject areas

  • Amino Acid Sequence
  • Antibodies, Monoclonal
  • B-Lymphocytes
  • Base Sequence
  • Cervix Uteri
  • Female
  • HIV Antibodies
  • HIV-1
  • Humans
  • Immunoglobulin Variable Region
  • Kenya
  • Leukocytes, Mononuclear
  • Molecular Sequence Data
  • Recombinant Proteins
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Identity

International Standard Serial Number (ISSN)

  • 0272-457X

Digital Object Identifier (DOI)

  • 10.1089/153685903321948021

PubMed ID

  • 12831535
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Additional Document Info

start page

  • 97

end page

  • 108

volume

  • 22

issue

  • 2

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