Biochemical estimation of hepatitis B surface antigen in recombinant yeast.
Purified recombinant hepatitis B surface antigen separated on polyacrylamide gels in the presence of sodium dodecyl sulphate has a very low staining index with Coomassie blue relative to a number of standard proteins. In contrast the protein stains better than average with silver nitrate. This property has been used to develop a semi-quantitative method of estimation of recombinant surface antigen in extracts of Saccharomyces cerevisiae producing this protein. The method can be used to follow purification protocols. It is quick, simple and since it measures the surface antigen biochemically, is independent of the aggregation state or conformation of the protein, a factor which can affect enzyme-linked immunoassays which rely on antigen-antibody interactions.
Wingfield
P T
PT
Department of Protein Chemistry, Biogen S.A., Carouge/Geneva, Switzerland.
Graber
P
P
Payton
M A
MA
D000135:Acid Phosphatase / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D012394:Rosaniline Dyes; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D012835:Silver Nitrate
D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue; D012835:Silver Nitrate; D000135:Acid Phosphatase