Purification of Glycoproteins by Selective Transport Using Concanavalin-Mediated Reverse Micellar Extraction

A novel methodology for coupling liquid‐liquid extraction with affinity interaction has been developed to selectively and efficiently purify and separate glycoproteins. The basis for the separation is the selective extraction of glycoproteins from an aqueous solution into a reverse micellar organic...

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Bibliographic Details
Published in:Biotechnology progress 1991-07, Vol.7 (4), p.330-334
Main Authors: Paradkar, Vikram M., Dordick, Jonathan S.
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Chemistry Techniques, Analytical - methods
Concanavalin A - metabolism
Dioctyl Sulfosuccinic Acid
Electrochemistry - methods
Fundamental and applied biological sciences. Psychology
Glycoproteins - isolation & purification
Glycoproteins - metabolism
Horseradish Peroxidase - metabolism
Kinetics
Methods. Procedures. Technologies
Micelles
Others
Surface-Active Agents
Various methods and equipments
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Summary:A novel methodology for coupling liquid‐liquid extraction with affinity interaction has been developed to selectively and efficiently purify and separate glycoproteins. The basis for the separation is the selective extraction of glycoproteins from an aqueous solution into a reverse micellar organic phase by using concanavalin A (a sugar‐binding lectin) as a facilitative carrier. Specifically, horseradish peroxidase (a common glycoprotein) can be bound to concanavalin A in an aqueous phase and then extracted into an AOT‐isooctane organic phase with negligible loss in enzyme activity. Virtually no extraction of peroxidase occurs in the absence of concanavalin A. Electron spin resonance studies have shown that the large lectin‐glycoprotein complex (96 000 daltons) resides in a nonaqueous environment within the reverse micelle, perhaps at the surfactant, water‐pool interface; hence, extraction of the large complex is feasible. The facilitative extraction has been extended to selective transport of peroxidase from a mixture of peroxidase and alkaline phosphatase (a nonglycosylated protein). This results in an efficient separation strategy with a separation factor of 16.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp00010a007