GlaxoSmithKline, Philadelphia, PA, USAM.
Cytotechnology. 2002 Jan;38(1-3):37-41. doi: 10.1023/A:1021189628274.
Baculovirus containing the mammalianCMV promoter, in place of the insect polyhedronpromoter (BacMam), has been used to transientlytransfect COS, CHO and CHOE1a (CHO cells expressing theE1a transcriptional activator). Using this system forthe expression of a cellular adhesion factor (SAF-3) Fcfusion protein in CHOE1a, we found that levels ofexpression were highest with a MOI of 100, 20mM sodiumbutyrate, at 34 degrees C. Production increased furtherif the cells were resuspended in fresh medium, about3 x 10(6) cells ml(-1), prior to addition of the virus. These conditions were used to express 3 secretedproteins, SAF-3-Fc, CD40-hexa his and Asp 2-Fc, and, at2 to 6 days post infection, protein levels ranged from4 ug ml(-1) to 25 ug ml(-1). Based on these results, theBacMam system represents a viable technique forproducing protein at ug ml(-1) levels in a relatively shortperiod of time.
含有哺乳动物 CMV 启动子的杆状病毒取代了昆虫多角体启动子（BacMam），已被用于瞬时转染 COS、CHO 和 CHOE1a（表达 E1a 转录激活剂的 CHO 细胞）。使用该系统在 CHOE1a 中表达细胞黏附因子（SAF-3）Fc 融合蛋白，我们发现 MOI 为 100、20mM 丁酸钠和 34°C 时表达水平最高。如果在添加病毒之前将细胞重新悬浮在新鲜培养基中（约 3 x 10(6)个细胞/ml），则产量会进一步提高。这些条件用于表达 3 种分泌蛋白，SAF-3-Fc、CD40-hexa his 和 Asp 2-Fc，在感染后 2 至 6 天，蛋白水平范围从 4ug/ml 到 25ug/ml。基于这些结果，BacMam 系统代表了一种可行的技术，可以在相对较短的时间内以 ug/ml 水平生产蛋白质。
