Suntara Fueangfung - Synthetic 5′-phosphorylated oligodeoxynucleotide purification through catching full-length sequences by polymerization

Version 1

      Publication Details (including relevant citation   information):

      RSC Advances., 2012,   2, 2803-2808

      DOI:  10.1039/C2RA01357F

      Abstract:

      The readily scalable catching by polymerization purification   technology has been further advanced to purify 5′-phosphorylated   synthetic oligodeoxynucleotides (ODNs). The new technology   utilizes a phosphoramidite that contains a fluoride-cleavable   diisopropylsilyl acetal linker and a polymerizable methacrylamide   group, and is capable of phosphorylation of ODN. For   purification, the phosphoramidite was coupled to the 5′-end of   full-length ODN on a synthesizer. Because failure sequences were   capped in each synthetic cycle, only the full-length sequences   were phosphinylated and acrylated. After cleavage and   deprotection, the crude ODN was subjected to polymerization under   typical acrylamide gel formation conditions. The full-length ODN   was incorporated into polymer. The failure sequences and other   impurities were simply removed by washing with water. Pure   full-length ODN that contained a 5′-phosphate group was cleaved   from the polymer with HF–pyridine. Reversed-phase (RP) HPLC   showed that the ODN was pure, and the recovery yield was higher   than that of typical preparative HPLC purification.

      Address (URL): http://pubs.rsc.org/en/content/articlelanding/2012/ra/c2ra01357f