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Methods of Analyzing Cell Membranes

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 Membrane proteins (MP's) are rapidly purified (~5 min) with a common table centrifuge.  This is achieved by precipitating MP's embedded in their native membranes thus, preserving protein functionality and circumventing the need for ultra-centrifuges! The method is conceptually new as it relies on a specific mechanism capable of conjugating phospholipid bilayers, regardless of their lipid & membrane protein composition. 
 The technology could also serve: Proteomics studies, as it allows fast and efficient separation of water-soluble proteins from MP's. Accordingly, samples containing mixtures of soluble and membrane proteins could be separated and applied on 2D distinct gels: one will primarily represent the MP's population whereas the other, primarily the water-soluble proteins. Following this simple protocol will substantially decrease the number of spots per gel, which in turn: (a) will simplify analysis (b) allow eluting target spots with minimum of protein impurities and (c) can be integrated into a high throughput mode and screening systems.    
 The method is the result of a long collaboration between three groups from three institutions: The Weizmann Institute of Science (Israel); The Indian Institute of technology, Bombay (India) and Ariel University (Israel).
 The abundant number and various types of proteins, coupled with the continued discovery of new proteins, means that researchers must continue to study their structure, function, modifications, and interactions. The need for protein characterization and functional studies drives basic research.
 Moreover, proteins are a major focus area for the pharmaceutical and diagnostics industry. Applications include protein-protein interaction studies, drug screening, target identification, biomarker discovery, protein therapeutics and diagnostic tests.
 In 2010, an estimated 22.3 million protein-purification reactions were conducted, 54.3% by tagged methods and 45.7% by non-tagged methods. Revenue for the global protein purification reagents market includes all reagents, consumables, and kits sold for tagged and non-tagged chromatography-based purification of all protein types globally. In 2010, this market earned $341.7 million in revenue at 4.8% growth.  
 Membrane-conjugation is achieved by the formation of strong complexes between hydrophobic chelators embedded at the membrane/water interface and metal ions (serving as mediators) in the aqueous phase, capable of binding several chelators simultaneously. Accordingly, membrane conjugation is triggered only upon addition of appropriate metals (e.g. Fe2+, Zn2+). Importantly, there is neither need for pH, temperature or ionic strength alterations nor any precipitants (e.g. PEGs, ammonium sulfate) and, membrane proteins are protected from denaturation during their purification as they are precipitated in their native membranous environment. 
 This single step process removes more than 85% of the impurity background with relatively good recovery yields (74-89%) of the targets studied (i.e. Bacteriorhodopsin, and Halorhodopsin). The fast protocol (~5 min.) which we had developed and the utilization of only a table centrifuge (rather than an ultracentrifuge) represent fundamental advantages, inherent to the technology.            
 Ready for marketing.   
 Dr. Patchornik received his B.Sc. and M.A degrees in Chemistry from Bar-Ilan University. He earned his Ph.D. in Biochemistry from the Weizmann Institute of Science and continued to a Postdoc. at UCLA. Upon his return to Israel, he co-founded a Biotech company (Affisink Biotechnology, Israel) and served as its CTO for five years. Since 2009, Dr. Patchornik is a faculty member in the Chemistry department at Ariel University where he studies alternative purification\crystallization methods for MP's.