Development and applications of surface-linked single chain antibodies against T-cell antigens

J Immunol Methods. 2001 Feb 1;248(1-2):77-90. doi: 10.1016/s0022-1759(00)00344-6.


In this report the use of surface-linkage to expand the potential experimental and therapeutic applications of single chain antibody (scFv) constructs is reviewed. A strategy for the generation and functional characterization of surface-linked scFvs that bind selectively to the T-cell proteins CD3epsilon, CD28, and CD152 (CTLA-4) is described in detail. Experimental examples are provided of the use of these constructs to study the positive and negative regulation of T-cell activation and to manipulate the in vivo immunogenicity of tumor cells. In addition, a novel system for Simultaneous T-cell Activation and Retroviral Transduction (START) is described in which retroviral packaging cells are rendered mitogenic for T lymphocytes by combined expression of surface-linked scFvs. Finally, the use of random mutagenesis and yeast surface display to increase the affinity and functional efficacy of scFv constructs is demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • Abatacept
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / immunology
  • Antigens, CD
  • Antigens, Differentiation / immunology*
  • CD28 Antigens / immunology*
  • CD3 Complex*
  • CTLA-4 Antigen
  • Humans
  • Immunoconjugates*
  • Immunoglobulin Fragments / genetics*
  • Immunoglobulin Fragments / immunology
  • Lymphocyte Activation
  • Molecular Sequence Data
  • Protein Engineering*
  • Receptors, Antigen, T-Cell / immunology*
  • Retroviridae / genetics
  • T-Lymphocytes / immunology*


  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Differentiation
  • CD28 Antigens
  • CD3 Complex
  • CD3E protein, human
  • CTLA-4 Antigen
  • CTLA4 protein, human
  • Immunoconjugates
  • Immunoglobulin Fragments
  • Receptors, Antigen, T-Cell
  • immunoglobulin Fv
  • Abatacept