N-terminally His-tagged hepatitis B core antigens: construction, expression, purification and antigenicity.
The core antigen of the hepatitis B virus (HBcAg) has been used widely as a diagnostic reagent for the identification of the viral infection. However, purification using the conventional sucrose density gradient ultracentrifugation is time consuming and costly. To overcome this, HBcAg particles displaying His-tag on their surface were constructed and produced in Escherichia coli. The recombinant His-tagged HBcAgs were purified using immobilized metal affinity chromatography. Transmission electron microscopy and enzyme-linked immunosorbent assay (ELISA) revealed that the displayed His-tag did not impair the formation of the core particles and the antigenicity of HBcAg.
Yap
Wei Boon
WB
Department of Microbiology, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Tey
Beng Ti
BT
Ng
Michelle Y T
MY
Ong
Swee Tin
ST
Tan
Wen Siang
WS
D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006512:Hepatitis B Core Antigens / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D046529:Microscopy, Electron, Transmission; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D022701:Virosomes / Q000648:ultrastructure
D006512:Hepatitis B Core Antigens; D011993:Recombinant Fusion Proteins; D022701:Virosomes