title: Development of an aptamer-affinity chromatography for efficient single step purification of Concanavalin A from Canavalia ensiformis.
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 41ntssDNA 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 ≥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 13mm, 20mm and 25mm diameter columns having a bed height of 60mm 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.
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi 110 007, India; Academy of Scientific and Innovative Research, CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi 110 007, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Ahirwar
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi 110 007, India; Academy of Scientific and Innovative Research, CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi 110 007, India. Electronic address: pnahar@igib.res.in.
authors.forename: Pradip
authors.initials: P
authors.lastname: Nahar
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism; D032449:Canavalia / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012965:Sodium Chloride
chemical_list: D052157:Aptamers, Nucleotide; D003208:Concanavalin A; D012965:Sodium Chloride
keywords: Aptamer-affinity chromatography; Canavalia ensiformis; Concanavalin A; DNA aptamer; NHS-activated agarose