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Technical and biological issues relevant to cell typing with aptamers.
Li, Na; Ebright, Jessica N; Stovall, Gwendolyn M; Chen, Xi; Nguyen, Hong Hanh; Singh, Amrita; Syrett, Angel; Ellington, Andrew D.
Afiliação
  • Li N; Department of Chemistry and Biochemistry, Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.
J Proteome Res ; 8(5): 2438-48, 2009 May.
Article em En | MEDLINE | ID: mdl-19271740
ABSTRACT
A number of aptamers have been selected against cell surface biomarkers or against eukaryotic tissue culture cells themselves. To determine the general utility of aptamers for assessing the cell surface proteome, we developed a standardized flow cytometry assay and carried out a comprehensive study with 7 different aptamers and 14 different cell lines. By examining how aptamers performed with a variety of cell lines, we identified difficulties in using aptamers for cell typing. While there are some aptamers that show excellent correlation between cell surface binding and the expression of a biomarker on the cell surface, other aptamers showed nonspecific binding by flow cytometry. For example, it has recently been claimed that an anti-PTK7 (protein tyrosine kinase 7) aptamer identified a new biomarker for leukemia cells, but data with the additional cell lines shows that it is possible that the aptamer instead identifies a propensity for adherence. Better understanding and controlling for the role of background and nonspecific binding to cells should open the way to using arrays of aptamers for describing and quantifying the cell surface proteome.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA de Cadeia Simples / Proteômica / Aptâmeros de Nucleotídeos / Proteínas de Membrana Limite: Humans Idioma: En Ano de publicação: 2009 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA de Cadeia Simples / Proteômica / Aptâmeros de Nucleotídeos / Proteínas de Membrana Limite: Humans Idioma: En Ano de publicação: 2009 Tipo de documento: Article