{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T19:23:01Z","timestamp":1777576981941,"version":"3.51.4"},"reference-count":28,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2010,12,15]],"date-time":"2010-12-15T00:00:00Z","timestamp":1292371200000},"content-version":"vor","delay-in-days":44,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Progress"],"published-print":{"date-parts":[[2010,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS\u2010CoV) is important for vaccine development. S<jats:sub>TR2<\/jats:sub> (an 88 kDa truncated SARS\u2010CoV TW1 S protein carrying the S fragments S\u201074\u2010253, S\u2010294\u2010739, and S\u20101129\u20101255) is capable of expressing a major form of glycoprotein as endo H\u2010sensitive (\u223c115 kDa) in CHO cells. To establish stable expressing cell clones, we transfected CHO\/dhFr\u2010cells with the amplifiable vectors ISID (IRES\u2010driven dhfr) and ISIZ (SV40\u2010driven dhfr) to select stepwise MTX, and observed enhanced \u223c115 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\u2010driven <jats:italic>dhfr<\/jats:italic> promoter generates greater gene amplification, which in turn enhances S<jats:sub>TR2<\/jats:sub> expression. Our results indicate that the \u223c115 kDa glycoform of S<jats:sub>TR2<\/jats:sub> protein was capable of increasing after gene amplification. The S<jats:sub>TR2<\/jats:sub> glycoform did not change between suspension and serum\u2010free cultures, suggesting that the stable and amplified cell clones analyzed in this study have potential for producing homologous S<jats:sub>TR2<\/jats:sub> on a large scale. \u00a9 2010 American Institute of Chemical Engineers Biotechnol. 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