[The purification of HBV full-length PreS protein in Pichia pastoris].
The Pichia pastoris strain GS115-PreS could produce a high expression level of full-length PreS protein that secreted to the supernatant after methanol induction in the fermentation. The Western blot analysis showed a single band with expected molecular mass of 48kD and that the major component of the particles was the full-length PreS protein (PreS1 + PreS2 + S) and small envelope protein (S) of 48 and 28 kD, respectively. Electron microscopy image showed PreS particles with 30 nm in diameter. The supernatants of the fermentation were desalted and concentrated. Purified PreS protein was obtained by DEAE-SFF anion exchange column chromatography and the PreS particles were obtained by ultracentrifugation and sucrose density gradient. The ELISA assay results proved that both full-length PreS protein and particles showed high immunogenicity and specificity. P/N ratio further demonstrated that the immunogenicity of the particles is higher than the full-length PreS protein.
Han
Xue
X
National Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
Ye
Lin-Bai
LB
Li
Bao-Zong
BZ
She
Ying-Long
YL
Ye
Li
L
Zheng
Hong
H
Gao
Bo
B
Gao
Jin-Rong
JR
Wu
Zheng-Hui
ZH
D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics
D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C050806:presurface protein 1, hepatitis B surface antigen; C050807:presurface protein 2, hepatitis B surface antigen