title: Ligand-guided selection of aptamers against T-cell Receptor-cluster of differentiation 3 (TCR-CD3) expressed on Jurkat.E6 cells.
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.
authors: 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.
authors: Hasan E
authors: HE
authors: Zumrut
authors: Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA.
authors: Mst N
authors: MN
authors: Ara
authors: Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA.
authors: George E
authors: GE
authors: Maio
authors: Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA.
authors: Nabeela A
authors: NA
authors: Van
authors: Department of Chemistry, Lehman College, City University of New York, 250 Bedford Park Blvd. West, Bronx, NY 10468, USA.
authors: Sana
authors: S
authors: Batool
authors: 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.
authors: Prabodhika R
authors: PR
authors: Mallikaratchy
mesh_terms: D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry / Q000276:immunology; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D017252:CD3 Complex / Q000737:chemistry* / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D019169:Jurkat Cells; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry* / Q000276:immunology; D052156:SELEX Aptamer Technique / Q000379:methods
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D052157:Aptamers, Nucleotide; D017252:CD3 Complex; D011948:Receptors, Antigen, T-Cell
keywords: Aptamer; CD3ε; Evolution; Ligand; Monoclonal antibody; T-cell Receptor