Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7617141 | Journal of Chromatography B | 2015 | 5 Pages |
Abstract
Herein, an aptamer-based affinity chromatography method for rapid and single step purification of Concanavalin A is developed and validated. We have used a 41 nt ssDNA aptamer of Con A (Con A aptabody) as an affinity reagent in the developed aptamer-affinity chromatography. Stationary phase of the method consists of surface functionalized agarose beads carrying covalently immobilized Con A-aptabody. Affinity purification of Con A from jack bean (Canavalia ensiformis) seed using developed aptamer-affinity columns has resulted in â¥66% recovery with 90% purity and 336-fold purification of Con A. The developed aptamer-affinity chromatography has shown efficient scalability and consistent purification when analysed over 13 mm, 20 mm and 25 mm diameter columns having a bed height of 60 mm each. Also, the developed aptamer-agarose columns were found to be reusable with recovery decrease of 12.9% in seven sequential cycles of purification. Therefore, the developed aptamer-affinity chromatography provides a novel, efficient and single-step methodology for isolation and purification of Con A.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Rajesh Ahirwar, Pradip Nahar,