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Phosphotungstate as a useful eluent for hepatitis‐B virus surface antigen purification by heparin‐sepharose affinity chromatography
Author(s) -
Tajima Yutaka,
Nagumo Fumio,
Nishimura Tadataka,
Tadano Jutaro
Publication year - 1992
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130060206
Subject(s) - hbsag , chemistry , elution , chromatography , affinity chromatography , heparin , sepharose , hepatitis b virus , antigen , column chromatography , virus , virology , biochemistry , enzyme , immunology , biology
On the basis of the heparin‐like effect of phosphotungstate (PTA), we have shown that it is useful for the purification of hepatitis‐B virus surface antigen (HBsAg) using heparin‐Sepharose affinity chromatography. HBsAg was eluted with 0.2–0.6 M NaCl. HBsAg was also eluted with PTA at approximately 1 mM, and the HBsAg fraction thus obtained contained fewer impurities than the corresponding fraction eluted with NaCl. Moreover, PTA yielded HBsAg and hepatitis‐B virus e‐antigen simultaneously. PTA may specifically reduce the affinity of HBsAg for heparin as well as simply competing with heparin for an anion‐binding site of HBsAg. Residual PTA in the eluate was easily decomposed by alkalization, which was useful for subsequent studies.