國家衛生研究院 NHRI:Item 3990099045/5309
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    题名: Generating stable chinese hamster ovary cell clones to produce a truncated SARS-CoV spike protein for vaccine development
    作者: Lin, SC;Leng, CH;Wu, SC
    贡献者: Vaccine Research and Development Center
    摘要: The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) is important for vaccine development. S<sub>TR2</sub> (an 88 kDa truncated SARS-CoV TW1 S protein carrying the S fragments S-74-253, S-294-739, and S-1129-1255) is capable of expressing a major form of glycoprotein as endo H-sensitive (~115 kDa) in CHO cells. To establish stable expressing cell clones, we transfected CHO/dhFr-cells with the amplifiable vectors ISID (IRES-driven dhfr) and ISIZ (SV40-driven dhfr) to select stepwise MTX, and observed enhanced ~115 kDa glycoform generation through gene amplification. Following stepwise MTX selection, we compared gene amplification levels between two vectors in engineered CHO cell chromosomes. These results confirm that the IRES-driven dhfr promoter generates greater gene amplification, which in turn enhancesSTR2 expression. Our results indicate that the ~115 kDa glycoform of STR2 protein was capable of increasing after gene amplification. The STR2 glycoform did not change between suspension and serum-free cultures, suggesting that the stable and amplified cell clones analyzed in this study have potential for producing homologous S<sub>TR2</sub> on a large scale.
    日期: 2010-11
    關聯: Biotechnology Progress. 2010 Nov;26(6):1733-1740.
    Link to: http://dx.doi.org/10.1002/btpr.480
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=8756-7938&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000285394700026
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=78650202713
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