title: Aptamer-containing surfaces for selective capture of CD4 expressing cells.
abstract: Aptamers have recently emerged as an excellent alternative to antibodies because of their inherent stability and ease of modification. In this paper, we describe the development of an aptamer-based surface for capture of cells expressing CD4 antigen. The glass or silicon surfaces were modified with amine-terminated silanes and then modified with thiolated RNA aptamer against CD4. Modification of the surface was first characterized by ellipsometry to demonstrate assembly of biointerface components and to show specific capture of recombinant CD4 protein. Subsequently, surfaces were challenged with model lymphocytes (cell lines) that were either positive or negative for CD4 antigen. Our experiments show that aptamer-functionalized surfaces have similar capture efficiency to substrates containing anti-CD4 antibody. To mimick capture of specific T-cells from a complex cell mixture, aptamer-modified surfaces were exposed to binary mixtures containing Molt-3 cells (CD4+) spiked into Daudi B cells (CD4-). 94% purity of CD4 cells was observed on aptamer-containing surfaces from an initial fraction of 15% of CD4. Given the importance of CD4 cell enumeration in HIV/AIDS diagnosis and monitoring, aptamer-based devices may offer an opportunity for novel cell detection strategies and may yield more robust and less expensive blood analysis devices in the future.
authors.affiliation: Department of Biomedical Engineering, University of California, Davis, California 95616, United States.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhou
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Dong-Sik
authors.initials: DS
authors.lastname: Shin
authors.forename: Jaime
authors.initials: J
authors.lastname: Silangcruz
authors.forename: Nazgul
authors.initials: N
authors.lastname: Tuleuova
authors.forename: Alexander
authors.initials: A
authors.lastname: Revzin
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology*; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D005898:Glass / Q000737:chemistry; D006801:Humans; D012821:Silanes / Q000737:chemistry; D012825:Silicon / Q000737:chemistry; D013499:Surface Properties; D013535:Suspensions
chemical_list: D052157:Aptamers, Nucleotide; D012821:Silanes; D013535:Suspensions; D012825:Silicon