Secretion of Particles of Hepatitis B Surface Antigen from Insect Cells Using a Baculovirus Vector

Kang, C. Yong; Bishop, David H. L.; Seo, Jin-Ho; Matsuura, Yoshiharu; Choe, Mu Hyeon

doi:10.1099/0022-1317-68-10-2607

Cited by 36 publications

(15 citation statements)

References 14 publications

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“…Efficient secretion of HBsAg particles into the cell supernatant was also reported for mammalian cells (Marion et al 1979;Liu et al 1982), but not for yeast (Valenzuela et al 1982;Hitzeman et al 1983;Janowicz et al 1991) or Lepidoptera cells upon recombinant baculovirus infection (Lanford et al 1989). Our findings in terms of HBsAg synthesis were comparable to those reported for Lepidoptera cells using the baculovirus expression system (Kang et al 1987;Lanford et al 1989) or mouse cells transfected using a bovine papilloma virus vector system (Hsiung et al 1984). Our approach allowed us to obtain HBsAg expression levels comparable to those previously published (Deml et al 1999).…”

Section: Discussionsupporting

confidence: 78%

Expression of the hepatitis B virus surface antigen in Drosophila S2 cells

et al. 2008

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Drosophila melanogaster S2 cells were transfected with a plasmid vector (pAcHBsAgHy) containing the S gene, coding for the hepatitis B virus surface antigen (HBsAg), under control of the constitutive drosophila actin promoter (pAc), and the hygromycin B (Hy) selection gene. The vector was introduced into Schneider 2 (S2) Drosophila cells by DNA transfection and a cell population (S2AcHBsAgHy) was selected by its resistance to hygromycin B. The pAcHBsAgHy vector integrated in transfected S2 cell genome and approximately 1,000 copies per cell were found in a higher HBsAg producer cell subpopulation. The HBsAg production varied in different subpopulations, but did not when a given subpopulation was cultivated in different culture flasks. Higher HBsAg expression was found in S2AcHBsAgHy cells cultivated in Insect Xpress medium (13.5 lg/1E7 cells) and SFX medium (7 lg/1E7 cells) in comparison to SF900II medium (0.6 lg/1E7 cells). An increase of HBsAg was observed in culture maintained under hygromycin selection pressure. Data presented in the paper show that S2AcHBsAgHy cells produce efficiently the HBsAg which is mainly found in the cell supernatant, suggesting that HBsAg is secreted from the cells. The data also show that our approach using the Drosophila expression system is suitable for the preparation of other viral protein preparation.

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Section: Discussionsupporting

confidence: 78%

Expression of the hepatitis B virus surface antigen in Drosophila S2 cells

et al. 2008

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“…Lower levels of expression have been reported for vectors that do not have all of the upstream sequences of the polyhedrin promoter (Matsuura et al, 1986(Matsuura et al, , 1987. Low level expression has been observed for certain gene products irrespective of the vector used (including the HBsAg, Kang et al, 1987;see below); the reason for the lower expression is not known.…”

Section: Introductionmentioning

confidence: 94%

“…35 mm dishes), the level of HBsAg synthesis by the recombinant baculovirus HBsYK 14 (Kang et al, 1987) has been reported to be equivalent to 1.5 mg per 109 infected cells. In large culture volumes (100 to 1000 ml), expression levels of the order of 0.3 to 0-5 mg per 109 infected ceils are obtained with the same virus.…”

Section: Recombinant Baculoviruses That Express Hbcag or Hbpcag Or Hbsagmentioning

confidence: 99%

“…In order to improve this, the HB S gene was transferred from the pAcRP6 vector to the pAcYM1 vector since higher expression has been observed with the latter for a variety of gene products (see Matsuura et al, 1987). In an initial study, the HBsAg gene was recovered from the transfer vector pAcRP6-HBsYK14 (Kang et al, 1987) by BamHI digestion and inserted directly into the BamHI site of transfer vector pAcYM1. The derived transfer vector was used to prepare a recombinant virus and the level of HBsAg synthesis was measured.…”

Section: Recombinant Baculoviruses That Express Hbcag or Hbpcag Or Hbsagmentioning

confidence: 99%

“…1). The entire S gene of HB virus was prepared by AcclI digestion of plasmid pAcRP6-HBsYKI4 (Kang et al, 1987), ligated into the BamHI site of pAcYM1, then digested with BamHI. The resulting 725 bp DNA fragment containing the S gene was inserted into the BamHI site of the vector pAcYM 1 to construct the recombinant transfer vector pAcYM1KTs (Fig.…”

Section: Construction Of Recombinant Transfer Vectors For Single Genementioning

confidence: 99%

See 2 more Smart Citations

Co-expression of the Hepatitis B Surface and Core Antigens Using Baculovirus Multiple Expression Vectors

Takehara

Ireland²,

Bishop

1988

Journal of General Virology

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SUMMARYThe hepatitis B (HB) virus DNA sequences coding for the pre-core (preC) or C antigens (HBpcAg, HBcAg) have been inserted into the baculovirus plasmid transfer vector, pAcYM1, such that the HB viral sequences are under the control of the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). Spodopterafrugiperda cells infected with either of the derived recombinant plasmids in the presence of infectious AcNPV DNA yielded recombinant, polyhedrin-negative viruses that expressed high levels of the respective HBpcAg or HBcAg (representing approx. 5 to 10~ and approx. 40~ of the stained cellular proteins, respectively). The particulate 27 nm HBcAgs have been purified to homogeneity from infected cell extracts by density gradient centrifugation. Dual expression transfer vectors containing the HBcAg gene sequences and the coding sequences of the HB viral S antigen (HBsAg), each gene under the control of its own copy of the polyhedrin promoter, have also been constructed and used to derive recombinant viruses. The recombinant with the HB C and S genes expressed high levels of the HBcAg (approx.

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Key issues in the selection of an expression system for vaccine antigens

Ellis

Gerety

1990

Journal of Medical Virology

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Three criteria by which the appropriate host cell is chosen for the expression of a recombinant-derived vaccine antigen are efficacy, safety, and scale-up. Efficacy for a vaccine antigen refers to the ability of the host cell to produce a vaccine antigen capable of eliciting a protective immune response. A concern for safety of a vaccine antigen relates to residual DNA in the final product, especially when derived from continuous mammalian cell lines as opposed to microbial cells. Since tens (or hundreds) of millions of doses of a widely used vaccine might be injected into healthy infants and young children during the lifetime of the product, safety is a critical issue, such that the use of a microbial expression system might be preferable to the use of a continuous cell line in certain circumstances.

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Secretion of Particles of Hepatitis B Surface Antigen from Insect Cells Using a Baculovirus Vector

Cited by 36 publications

References 14 publications

Expression of the hepatitis B virus surface antigen in Drosophila S2 cells

Expression of the hepatitis B virus surface antigen in Drosophila S2 cells

Co-expression of the Hepatitis B Surface and Core Antigens Using Baculovirus Multiple Expression Vectors

Key issues in the selection of an expression system for vaccine antigens

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