Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system

R. E. Landford, V. Luckow, R. C. Kennedy, G. R. Dreesman, L. Notvall, M. D. Summers

    Research output: Contribution to journalArticlepeer-review

    60 Scopus citations

    Abstract

    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.

    Original languageEnglish (US)
    Pages (from-to)1549-1557
    Number of pages9
    JournalJournal of virology
    Volume63
    Issue number4
    DOIs
    StatePublished - 1989

    ASJC Scopus subject areas

    • Microbiology
    • Immunology
    • Insect Science
    • Virology

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