[Purification of hepatitis B virus antigen and its application in detection of human serum antibody].
A recombinant plasmid DNA pBV220/eAg provided by professor Zhou Jian was transferred into DH5 alpha celes. The effectively expressed strain was harvested after cloning. It was then incubated in 30 degrees C for 3 hrs and 42 degrees C for 6 hrs, the HBeAg protein was induced. After purification of the protein by DEAE-Sepharose Fast Flow chromatography, the purified protein was analyzed by SDS-PAGE electrophoresis and Coomassic brilliant blue R250 stain and only one single protein band was seen, Its specificity was demonstrated by Western-blot and Immunodot. 49 HBeAb positive sera of hepatitis B patients could be specifically detected with the purified HBeAg by ELISA and immunostrip assay, but the latter was more sensitive and specific.
Zhu
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Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
Gu
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D000914:Antibodies, Viral / Q000097:blood*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D006509:Hepatitis B / Q000276:immunology / Q000821:virology; D006513:Hepatitis B e Antigens / Q000096:biosynthesis / Q000302:isolation & purification*; D006515:Hepatitis B virus / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D000914:Antibodies, Viral; D006513:Hepatitis B e Antigens; D011994:Recombinant Proteins