ISSN:
0173-0835
Keywords:
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
Free flow electrophoresis was used for the purification of human tissue plasminogen activator (t-PA) produced by recombinant yeast cells. This method was employed for three principal reasons: (a) yeast t-PA purified by immunoaffinity chromatography was not pure enough to carry out protein chemical characterization, (b) the amount of yeast t-PA available for purification was extremely limited, (c) t-PA has a strong tendency to adsorb to support material; a support-free separation method therefore seemed to be a reasonable way to further purify the enzyme. Free flow electrophoresis was first performed using solutions containing the soluble proteins of yeast cell extract, and secondly, using t-PA fractions prepared by immunoaffinity chromatography, Yeast t-PA purified by immunoaffinity chromatography and then by free flow electrophoresis is more than 80% pure and suitable for characterization experiments such as glycosylation studies or N-terminal sequence determination.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elps.1150070805
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