[Expression of hepatitis delta antigen Inner Mongolian strain in prokaryotic cell and analysis of its antigenicity].
OBJECTIVE
To obtain high yield and good antigenic activity of HDV L-Ag and to detect different regional patients' sera to test the purified antigen's antigenicity.


METHODS
Hepatitis delta virus' sequence was obtained from Inner Mongolian patient by using RT-PCR and PCR methods, PET43a was used and His-tag was added at the HDV L-Ag 5' and 3' to construct the recombinant expression plasmid, transform the plasmid into host bacterium BL21 and induce it with IPTG. The expression supernatant was purified by saturated (NH4)2SO4 and affinity chromatography. The activity and antigenicity of the expressed product were analyzed by using EIA.


RESULTS
Comparison of results obtained with detection by using the expressed protein coated plate and ABBOTT Murex anti-Delta (total) of 15 positive and 10 negative sera, the consistency was good (100%).


CONCLUSIONS
EIA proved that the purified antigen had good antigenicity, no serological difference was found in detection between different region's sera, therefore the purified delta antigen may be useful in diagnostic and other research.
Yang
Ying-chao
YC
Research Laboratory of Hepatitis, Institute for Viral Disease Control and Prevention, Chinese Center For Disease Control and Prevention, Beijing 100052, China.
Yi
Yao
Y
Zhao
Hong-lan
HL
Zhang
Wen-ying
WY
Chen
Si-yong
SY
Bi
Sheng-li
SL
D000595:Amino Acid Sequence; D002681:China; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D003699:Hepatitis D / Q000097:blood / Q000821:virology; D003698:Hepatitis Delta Virus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D035923:Hepatitis delta Antigens / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism
D035923:Hepatitis delta Antigens; D011994:Recombinant Proteins