Expression of recombinant herpes simplex virus type 2 glycoprotein D by high-density cell culture of <Emphasis Type="Italic">Spodoptera frugiperda</Emphasis> |
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Authors: | Tao Liu Ji-Feng Liu Hui-Jun Dong Wei Zheng Zhi-Cheng Huang Shui-Fen Zhu |
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Institution: | 1.Microbiology Laboratory,Hangzhou Center for Disease Control and Prevention,Hangzhou,People’s Republic of China;2.Department of Dermatology,The Third Hospital of Hangzhou,Hangzhou,People’s Republic of China;3.Lunan Pharmaceutical Group Corporation,Linyi,People’s Republic of China |
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Abstract: | Herpes simplex virus type 2 (HSV-2) is the major cause of genital herpes in humans. The glycoprotein D of HSV-2 (gD2) is a
promising subunit vaccine candidate for the treatment of genital herpes. The aim of the present study was to express a biologically
active recombinant gD2 in eukaryotic baculovirus system in quantities sufficient for further studies. Human cDNA encoding
a gD2 protein with 393 amino acids was subcloned into the pFastBac HTb vector and the recombinant protein was expressed in
Spodoptera frugiperda (Sf9) cells by high-density cell culture. In a stirred bioreactor, the key limiting factors including glucose concentration,
glutamine concentration and dissolved oxygen (DO) were optimized for high-density cell growth. The Sf9 cell density could
reach 9.6×106 cells/mL and the yield of recombinant gD2 protein was up to 192 mg/L in cell culture under the optimal conditions of 15 mM
glucose, 0.4 g/L glutamine and 40% DO. Production of significant amounts of pure, full-length gD2 opened up the possibility
to investigate novel functions of gD2. Moreover, the purified recombinant gD2 protein revealed a partial prophylactic immune
function in genital herpes of guinea pigs infected with HSV-2. |
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