期刊
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
卷 44, 期 7, 页码 725-737出版社
TAYLOR & FRANCIS INC
DOI: 10.1080/10826068.2013.868358
关键词
alpha(1)-antitrypsin; chromatography; Cohn fraction IV; mass spectrometry; purification
类别
资金
- P.R. China Sichuan Province Science and Technology Support Program [2009SZ0217]
Alpha(1)-antitrypsin is a kind of plasma protein that requires a sequence of different fractionation steps to get generally. To report an effective process for isolating and purifying alpha(1)-antitrypsin from Cohn Fraction IV based upon a new immunoaffinity chromatography medium, named Alpha-1 Antitrypsin Select,'' characterization of alpha(1)-antitrypsin (alpha 1-AT) was performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, and tandem mass spectroscopy. Total protein content was determined by the method of Bradford under visible light absorption at 595 nm. Pretreatment process and the immunoaffinity chromatography step achieved a 60.35 +/- 1.39% yield. Thus, an overall 71.68 +/- 1.32 fold increase in purity and a 41.88 +/- 6.98% yield were obtained from plasma. The alpha 1-AT had a specific activity of about 1.00-1.05 PU/mg. This technique will develop an effective process for isolating and purifying, with high purity and specific activity, alpha(1)-antitrypsin from Cohn Fraction IV or human whole plasma, which could be an efficient and scaled-up method for alpha(1)-antitrypsin products purification.
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