Preparative reversed-phase liquid chromatography of proteins from rabbit skeletal troponin, a multi-protein complex

Journal of Chromatography. a
C T Mant, R S Hodges

Abstract

A reversed-phase high-performance liquid chromatography protocol for purification of all proteins in a multi-protein (TnI, TnC, TnT, tropomyosin) complex from rabbit skeletal muscle has been developed, enabling efficient purification of sample amounts ranging from 43 mg of protein complex on a standard analytical column, to 1400 mg on a column of 21.2 mm I.D. and finally, to 5700 mg on a column of 50 mm I.D. Due to problems associated with scale-up procedures for these proteins (e.g. aggregation and/or solubility issues), an initial sample fractionation was devised whereby 50% of the TnC component was precipitated with acetonitrile prior to sample introduction on the RPLC column. By subsequently taking advantage of sample overload conditions to enhance the displacement effect between sample components, coupled with very slow gradient conditions (0.1% acetonitrile/min), we were able to achieve excellent protein separations at high yields of purified proteins.

References

Aug 1, 1992·Journal of Muscle Research and Cell Motility·Z GrabarekJ Gergely
Jun 15, 1992·Analytical Chemistry·J G DorseyH G Barth
Jul 12, 1991·Journal of Chromatography·R S HodgesC T Mant
Jan 1, 1987·Annual Review of Biophysics and Biophysical Chemistry·A S Zot, J D Potter
Jan 1, 1968·Progress in Biophysics and Molecular Biology·S Ebashi, M Endo
Feb 1, 1971·Journal of Biochemistry·S EbashiF Ebashi
Jan 1, 1983·Methods in Enzymology·F E Regnier
Jan 1, 1982·Methods in Enzymology·J D Potter
Jun 1, 1995·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·C S Farah, F C Reinach
Apr 28, 1995·Journal of Chromatography. a·K BárányC S Giometti
Jan 1, 1996·Methods in Enzymology·C T Mant, R S Hodges
Aug 14, 2001·Journal of Cellular Biochemistry·S M NgaiR S Hodges

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Citations

Jul 31, 2007·Journal of Chromatography. a·João Ezequiel OliveiraMaria Teresa Carvalho Pinto Ribela
Apr 24, 2012·Journal of Chromatography. a·Martina Srajer GajdosikDjuro Josic
Sep 4, 2010·Current Protocols in Protein Science·Djuro Josic, Spomenka Kovac
Jul 9, 2004·Journal of Biochemical and Biophysical Methods·Even MansethRagnar Flengsrud

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