Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system.
The baculovirus Autographa californica nuclear polyhedrosis virus was used as an expression vector to produce hepatitis B virus surface antigen with and without the pre-S domain. The S gene product was expressed as both fusion and nonfusion polypeptides. No difference was observed in the posttranslational modification of the fusion and nonfusion polypeptides. The S proteins were not secreted into the medium but were inserted into the endoplasmic reticulum, glycosylated, and partially extruded into the lumen of the endoplasmic reticulum as 22-nm lipoprotein particles. The oligosaccharide chains on the insect cell-derived S protein were of the N-linked high-mannose form, in contrast to the complex-type oligosaccharides detected on plasma-derived hepatitis B virus surface antigen. The pre-S-S polypeptides were inserted into the endoplasmic reticulum, glycosylated, and modified by fatty acid acylation with myristic acid. A procedure was developed to purify the S protein from cellular membranes by using detergent extraction and immunoaffinity chromatography. The purified S protein was in the form of protein-detergent micelles and was highly antigenic and immunogenic.
Lanford
R E
RE
Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas 78284.
Luckow
V
V
Kennedy
R C
RC
Dreesman
G R
GR
Notvall
L
L
Summers
M D
MD
D000818:Animals; D002460:Cell Line; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis; D006031:Glycosylation; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000276:immunology; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D009226:Myristates / Q000378:metabolism; D011499:Protein Processing, Post-Translational
D004274:DNA, Recombinant; D006023:Glycoproteins; D006514:Hepatitis B Surface Antigens; D009226:Myristates