Aptamer affinity chromatography: combinatorial chemistry applied to protein purification.
The systematic evolution of ligands by exponential enrichment process is a combinatorial chemistry method that allows the identification of specific oligonucleotide sequences, known as aptamers, that bind to a desired target molecule with high affinity and specificity. Here, a DNA-aptamer specific for human L-selectin was immobilized to a chromatography support to create an affinity column. This column was effectively applied as either the first or second step in the purification of a recombinant human L-selectin-Ig fusion protein from Chinese hamster ovary cell-conditioned medium. The fusion protein was efficiently bound to the column and efficiently eluted by gentle elution schemes. Application of the aptamer column as the initial purification step resulted in a 1500-fold purification with an 83% single step recovery. These results demonstrate that oligonucleotide aptamers can be effective affinity purification reagents.
Romig
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TS
NeXstar Pharmaceuticals, Boulder, CO 80301, USA.
Bell
C
C
Drolet
D W
DW
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006224:Cricetinae; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D019020:Directed Molecular Evolution; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D019041:L-Selectin / Q000737:chemistry / Q000302:isolation & purification*
D017931:DNA Primers; D019041:L-Selectin; D004247:DNA