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

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

Author information

1
Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA; Ph.D. Program in Biochemistry, The Graduate Center of the City University of New York, New York, NY 10016, USA.
2
Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA.
3
Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA; Ph.D. Program in Biochemistry, The Graduate Center of the City University of New York, New York, NY 10016, USA; Ph.D. Program in Chemistry, The Graduate Center of the City University of New York, New York, NY 10016, USA. Electronic address: prabodhika.mallikaratchy@lehman.cuny.edu.

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

PMID:
27519622
PMCID:
PMC5593316
DOI:
10.1016/j.ab.2016.08.007
[Indexed for MEDLINE]
Free PMC Article

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