Phosphotungstate as a useful eluent for hepatitis-B virus surface antigen purification by heparin-sepharose affinity chromatography.
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.
Tajima
Y
Y
Clinical Laboratory, Saga Medical School Hospital, Japan.
Nagumo
F
F
Nishimura
T
T
Tadano
J
J
D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006493:Heparin / Q000737:chemistry; D006514:Hepatitis B Surface Antigens / Q000302:isolation & purification*; D006801:Humans; D010772:Phosphotungstic Acid / Q000737:chemistry*
D006514:Hepatitis B Surface Antigens; D010772:Phosphotungstic Acid; D006493:Heparin