title: Automated solid-phase synthesis of high capacity oligo-dT cellulose for affinity purification of poly-A tagged biomolecules.
abstract: Affinity purification of poly-adenylated biomolecules using solid supports that are derivatized with poly-thymidine oligonucleotides provides a powerful method for isolating cellular mRNA. These systems have also been used to purify mRNA-peptide fusions generated by RNA-display. However, the commercial source for high capacity oligo-dT cellulose was recently discontinued. To overcome this problem, we have developed a low cost solid-phase synthesis protocol to generate oligo-dT cellulose. Comparative binding studies indicate that chemically synthesized oligo-dT cellulose functions with superior loading capacity when compared to the discontinued product. We suggest that this method could be used to synthesize oligo-dT resin for routine purification of poly-adenylated biomolecules.
authors.affiliation: The Biodesign Institute, Arizona State University, Tempe, AZ 85287-5301, USA.
authors.forename: Sujay P
authors.initials: SP
authors.lastname: Sau
authors.affiliation: The Biodesign Institute, Arizona State University, Tempe, AZ 85287-5301, USA.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Larsen
authors.affiliation: The Biodesign Institute, Arizona State University, Tempe, AZ 85287-5301, USA; Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287-5301, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: Chaput
mesh_terms: D002482:Cellulose / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism*; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D060327:Solid-Phase Synthesis Techniques / Q000379:methods*
chemical_list: D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; C027903:oligo (dT); D011061:Poly A; D002482:Cellulose
keywords: Affinity purification; Oligo-dT cellulose; mRNA display