Analysis and characterization of hepatitis B vaccine particles synthesized from Hansenula polymorpha.
The biochemical and physical properties of hepatitis B virus (HBV) small surface antigen (S-HBVsAg) from Berna Biotech Korea Corp. were systematically analyzed and characterized. Through various electrophoresis and immunoblotting assay of S-HBVsAg and its proteolytic products, it was confirmed that the S-HBVsAg vaccine particles are present in the form of covalent multimers that are assembled via strong intermolecular disulfide bonds. The S-HBVsAg particles contain no N-glycosylation moiety but some O-glycosidically linked mannoses. Evidently from N-terminus sequencing of both monomers and dimers that are formed by complete and partial reduction, respectively, of the S-HBVsAg particles under reducing SDS-PAGE condition, it is evident that each polypeptide within S-HBVsAg particles has authentic sequence of N-terminus. Denaturation plot shows that the S-HBVsAg vaccine particles were extremely stable especially in the solution with high acidity. This stability property of S-HBVsAg vaccine particles could provide very useful information for the optimization of the downstream process of recombinant S-HBVsAg particles synthesized from yeast cultures.
Seo
Hyuk-Seong
HS
Department of Chemical and Biological Engineering, Korea University, Anam-Dong 5-1, Sungbuk-Ku, Seoul 136-713, South Korea.
Park
Jin-Seung
JS
Han
Kyung-Yeon
KY
Bae
Kyung-Dong
KD
Ahn
Sang Jeom
SJ
Kang
Hyun Ah
HA
Lee
Jeewon
J
D015153:Blotting, Western; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D017325:Hepatitis B Vaccines / Q000096:biosynthesis* / Q000235:genetics; D046911:Macromolecular Substances; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D046911:Macromolecular Substances