Ligand-guided selection of aptamers against T-cell Receptor-cluster of differentiation 3 (TCR-CD3) expressed on Jurkat.E6 cells

Anal Biochem. 2016 Nov 1;512:1-7. doi: 10.1016/j.ab.2016.08.007. Epub 2016 Aug 9.

Abstract

We recently introduced a screening technology termed ligand-guided selection, (LIGS), to selectively identify target-specific aptamers from an evolved cell-SELEX library. Cell-SELEX utilizes a large combinatorial single-stranded oligonucleotide library and progressively selects DNA ligands against whole cells with variable DNA-binding affinities and specificities by repeated rounds of partition and amplification. LIGS exploits the partition step and introduces a secondary, pre-existing high-affinity monoclonal antibody (mAb) ligand to outcompete and elute specific aptamers towards the binding target of the antibody, not the cell. Here, using anti-CD3ε mAb against the cluster of differentiation 3 (CD3ε), as the guiding ligand against one of the domains of the T-cell Receptor (TCR) complex expressed on Jurkat.E6 cells, we discovered three specific aptamers against TCR complex expressed on an immortalized line of human T lymphocyte cells. In sum, we demonstrate that specific aptamers can be identified utilizing an antibody against a single domain of a multidomain protein complex in their endogenous state with neither post- nor pre-SELEX protein manipulation.

Keywords: Aptamer; CD3ε; Evolution; Ligand; Monoclonal antibody; T-cell Receptor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal, Murine-Derived / chemistry
  • Antibodies, Monoclonal, Murine-Derived / immunology
  • Aptamers, Nucleotide / chemistry*
  • CD3 Complex / chemistry*
  • CD3 Complex / immunology
  • Gene Expression*
  • Humans
  • Jurkat Cells
  • Receptors, Antigen, T-Cell / chemistry*
  • Receptors, Antigen, T-Cell / immunology
  • SELEX Aptamer Technique / methods

Substances

  • Antibodies, Monoclonal, Murine-Derived
  • Aptamers, Nucleotide
  • CD3 Complex
  • Receptors, Antigen, T-Cell