P. Bridonneau et al., Purification of a highly modified RNA-aptamer - Effect of complete denaturation during chromatography on product recovery and specific activity, J CHROMAT B, 726(1-2), 1999, pp. 237-247
To evaluate RNA-aptamers as potential drug candidates, efficient and scalea
ble purification protocols are needed. Because aptamers are highly structur
ed and rigid molecules, denaturation during the purification process is a c
ritical aspect to obtain a pure and active product. A two-step chromatograp
hic procedure was developed to purify a synthetic anti-VEGF aptamer at the
preparative scale. A reversed-phase chromatographic step was optimized with
a highly hydrophobic ion pairing reagent, followed by ion-exchange chromat
ography in which heat and a chaotropic salt were used. Because of the prese
nce of 2'-modified ribose, denaturation conditions had to be optimized in b
oth chromatographic steps to achieve a fully active molecule. (C) 1999 Else
vier Science B.V. All rights reserved.