SpringerProtocols: Abstract: Protein Separation Using Immobilized Phospholipid Chromatography

20. Protein Separation Using Immobilized Phospholipid Chromatography

By: Tzong-Hsien Lee², Marie-Isabel Aguilar²

Abstract

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The chromatographic support containing monolayers of phospholipids offers novel modes in analyzing and separating proteins. The polar choline headroups on immobilized phosphatidylcholine were used for the affinity purification of phospholipase A (PLA). The purification process involves removing the contaminating proteins with detergent additives to the elution buffer such as short-chain alkylsulfonates. The lipid-bound PLA was eluted with acetonitrile or octyllysophosphatidylcholine. The purity of PLA was approximately 70% based on densitometric scans ofel electrophoresis. These results suggest that the lipid-immobilized chromatography may be applied to develop purification methods for PLA, enzymes, and membrane proteins obtained from diverse cells.

Affiliation(s): (2) Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia

Book Title: Affinity Chromatography: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 421 | Pub. Date: Nov-30-2007 | Page Range: 295-302 | DOI: 10.1007/978-1-59745-582-4_20

Subject: Cell Biology

Key Words: Immobilized lipid chromatography - membrane proteins - detergent - organic solvent

References

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