Aptamer affinity chromatography:: combinatorial chemistry applied to protein purification |
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Authors: | Timothy S. Romig Carol Bell Daniel W. Drolet |
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Affiliation: | NeXstar Pharmaceuticals, 2860 Wilderness Place, Boulder, CO 80301, USA |
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Abstract: | 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 -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 -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. |
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Keywords: | Proteins -Selectin |
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