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 journalArticle

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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
StatePublished - 1989
Externally publishedYes

Fingerprint

Hepatitis B virus
Baculoviridae
Protein S
surface antigens
Hepatitis B Surface Antigens
Insects
polypeptides
Endoplasmic Reticulum
Peptides
insects
endoplasmic reticulum
Oligosaccharides
Detergents
proteins
oligosaccharides
detergents
cells
Nucleopolyhedrovirus
Acylation
Myristic Acid

ASJC Scopus subject areas

  • Immunology

Cite this

Landford, R. E., Luckow, V., Kennedy, R. C., Dreesman, G. R., Notvall, L., & Summers, M. D. (1989). Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system. Journal of Virology, 63(4), 1549-1557.

Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system. / Landford, R. E.; Luckow, V.; Kennedy, R. C.; Dreesman, G. R.; Notvall, L.; Summers, M. D.

In: Journal of Virology, Vol. 63, No. 4, 1989, p. 1549-1557.

Research output: Contribution to journalArticle

Landford, RE, Luckow, V, Kennedy, RC, Dreesman, GR, Notvall, L & Summers, MD 1989, 'Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system', Journal of Virology, vol. 63, no. 4, pp. 1549-1557.
Landford, R. E. ; Luckow, V. ; Kennedy, R. C. ; Dreesman, G. R. ; Notvall, L. ; Summers, M. D. / Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system. In: Journal of Virology. 1989 ; Vol. 63, No. 4. pp. 1549-1557.
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