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Adsorption and elution behaviors of bovine serum albumin in metal-chelated affinity cryogel beds

Paper ID Volume ID Publish Year Pages File Format Full-Text
4331 221 2008 6 PDF Available
Title
Adsorption and elution behaviors of bovine serum albumin in metal-chelated affinity cryogel beds
Abstract

Metal-chelated supermacroporous cryogels are effective supports for affinity chromatographic separation of biomolecules in downstream processes. In this work, polyacrylamide cryogel beds were prepared in glass columns (16 mm inner diameter) and coupled with iminodiacetic acid (IDA). These cryogels were loaded with Zn2+ and Ni2+ and the so-called Zn2+-IDA-cryogels and Ni2+-IDA-cryogels were obtained. Permeabilities and height equivalent to theoretical plates (HETPs) of these cryogel beds were measured and the cryogel structure was analyzed using scanning electron microscopy (SEM). Bovine serum albumin (BSA) was employed as a model protein to elucidate the adsorption and elution behaviors of these cryogels under various conditions, such as different flow rate, solution pH, and composition of the eluents. The results showed that the Zn2+-IDA-cryogels and Ni2+-IDA-cryogels in this study had interconnected supermacropores and high water permeabilities (∼10−11 m2). The loading flow velocity had a weak influence on the breakthrough curves and binding capacities for BSA, while the solution pH had an evident effect on the binding capacities for BSA in these cryogels. Maximum binding capacity for BSA was observed near the isoelectric point of BSA. The bound BSA can be eluted effectively using an imidazole solution. A low-eluting flow rate was found to be beneficial to the elution process. Possible mechanisms were proposed and discussed.

Keywords
Supermacroporous cryogels; Immobilized metal affinity chromatography; Bovine serum albumin; Protein adsorption; Binding capacity; Breakthrough; Elution
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Adsorption and elution behaviors of bovine serum albumin in metal-chelated affinity cryogel beds
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 42, Issue 3, 1 December 2008, Pages 237–242
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us