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Generation and characterization of a recombinant human CCR5-specific antibody. A phage display approach for rabbit antibody humanization

Academic Article
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Overview

authors

  • Steinberger, P.
  • Sutton, J. K.
  • Rader, Christoph
  • Elia, M.
  • Barbas III, Carlos

publication date

  • November 2000

journal

  • Journal of Biological Chemistry  Journal

abstract

  • We describe the isolation of a CCR5-specific antibody, ST6, from an antibody phage display library generated from an immune rabbit. ST6 was previously shown to efficiently prevent the surface expression of CCR5 when expressed intracellularly (Steinberger, P., Andris-Widhopf, J., Buhler, B., Torbett, B. E., and Barbas, C. F., III (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 805-810). Because ST6 has therapeutic potential in human immunodeficiency virus, type 1 disease, its humanization was desired to minimize the potential for immunogenicity. ST6 was humanized using a phage display-based approach. Like the parental rabbit clone, the humanized version ST6/34 efficiently prevented the surface expression of CCR5. The conserved linear peptide epitope bound by these antibodies was mapped using phage display. Both ST6 as well as the humanized anti-CCR5 antibody ST6/34 were produced as complete IgG antibodies and shown to bind to cell surface CCR5.

subject areas

  • Amino Acid Sequence
  • Animals
  • Antibodies
  • Antibody Specificity
  • Base Sequence
  • CCR5 Receptor Antagonists
  • Chemokines
  • Epitope Mapping
  • Genetic Therapy
  • HIV-1
  • Humans
  • Immunoglobulin Fab Fragments
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Molecular Sequence Data
  • Peptide Library
  • Protein Engineering
  • Rabbits
  • Receptors, CCR5
  • Receptors, HIV
  • Recombinant Fusion Proteins
  • Sequence Alignment
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Identity

International Standard Serial Number (ISSN)

  • 0021-9258

Digital Object Identifier (DOI)

  • 10.1074/jbc.M002765200

PubMed ID

  • 10969070
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Additional Document Info

start page

  • 36073

end page

  • 36078

volume

  • 275

issue

  • 46

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