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AFFINITY CHROMATOGRAPHY IN THE SEPARATION OF HUMAN ALPHA 1 ‐ANTITRYPSIN (α 1 ‐AT) AND ANTITHROMBIN‐III (AT‐III)
Author(s) -
D'Souza S,
Ananthakrishnan R
Publication year - 1979
Publication title -
australian journal of experimental biology and medical science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0004-945X
DOI - 10.1038/icb.1979.26
Subject(s) - antithrombin , chemistry , chromatography , sephadex , size exclusion chromatography , affinity chromatography , sepharose , ion exchange , ion chromatography , yield (engineering) , heparin , biochemistry , ion , enzyme , materials science , organic chemistry , metallurgy
Summary The two antiproteases α 1 ‐antitrypsin (α 1 ‐AT) and antithrombin‐III (AT‐III) have been purified simultaneously from human plasma. Purification procedure consisted of gel filtration on Sephadex G‐200 after initial processing of plasma, followed by ion exchange chromatography on DEAE‐Sephadex A50 and DEAE‐celllulose, at a pH of 9·0 and pH 8·3 respectively. The two proteins could not be separated by any of these procedures including a lower pH (7·4) in ion exchange chromatography. Affinity chromatography on heparin‐Sepharose separated the proteins since α 1 ‐AT did not bind to the matrix. Alpha 1 ‐AT unbound to the heparin‐Sepharose was subsequently purified through con A‐Sepharose affinity column. The final yield of both the proteins was about 20%. The molecular weight estimated on SDS electrophoresis for AT‐III and α 1 ‐AT was 63,000 and 50,000, respectively.