Resolution of Structurally Similar Glycoproteins by Affinity-BasedReversed Micellar Extraction and Separation
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Authors
Choe, J.H.
VanderNoot, V.A.
Linhardt, Robert J.
Dordick, J.S.
Issue Date
1998
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
Affinity-based reversed micellar extraction and separation (ARMES) has proven effective in separating glycoproteins from nonglycosylated proteins from natural sources. The ability of ARMES to resolve closely related glycoproteins is of paramount importance if ARMES is to be used in glycoform resolution. It is demonstrated that ARMES can resolve the structurally similar soybean peroxidase (SBP; MW 37 kDa, pI 4.1) and αt-acid glycoprotein (AGP; MW 43 kDa, pI 3.7), both of which have affinity for Concanavalin A (Con A) (the affinity ligand). SBP was almost exclusively extracted at pH 8 and above, with a separation factor greater than 50 (resolution ∼ 20), far better than was possible using Con A affinity chromatography (R ∼ 0.25, separation factor ∼ 2). Model calculations suggest that differences in affinity measured by an equilibrium-building assay cannot account for the favorable extraction of SBP over AGP at higher pH. Hydrophobic interactions and/or charge shielding appear to affect partitioning of the lectin - glycoprotein complexes and add greatly to the selectivity of extraction in ARMES, especially at higher pH values.
Description
AIChE Journal, 44, 2542-2548
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Resolution of Structurally Similar Glycoproteins by Affinity-BasedReversed Micellar Extraction and Separation, J.-H. Choe, V.A. VanderNoot,R.J. Linhardt, J.S. Dordick, AIChE Journal, 44, 2542-2548, 1998.