一株高水平表达重组蛋白昆虫细胞系的建立

Establishment of an insect cell line for high level expression of recombinant protein

报道了一株来自粉纹夜蛾Trichoplusiani脂肪体的传代细胞系 ,在辅以 5%胎牛血清的商品无血清培养基Excell 4 0 0中 ,细胞群体倍增时间为 2 2 9h ,最高密度可达 2 2× 10 6 mL ,该细胞对苜蓿丫纹夜蛾多粒包埋型多角体病毒 (AcMNPV)极为敏感 ,增殖AcMNPV多角体平均每个细胞达86个 ,表达由AcMNPV构建的重组蛋白的水平较高 ,β 半乳糖苷酶的表达水平为 ( 2 2 5 5± 13 4 )IU mL ;碱性磷酸酶的表达水平为 ( 4 7± 0 61)IU mL ,是一株高水平表达重组蛋白的传代细胞系 ,命名为HNU Tn FB1。

A new cell line was established from Trichoplusia ni fatbody in Excell 400 medium supplemented with 5% FBS. The maximum cell density reached 2 2×10 6 cells /mL and the population doubling time was 22 9 h for this cell line. The cell line was highly sensitive to AcMNPV, and produced 86 occlusion bodies per cell. The expression levels for two recombinant proteins, β galactosidase and alkaline phosphatase, in this cell line were compared to that in IPLB Sf 21 and BTI 5B1. The activities of β galactosidase and alkaline phosphatase from the novel cell line were (225 5±13 4)IU/mL and (4 7±0 61) IU/mL respectively. The expression levels from the fat body cells and from BTI TN 5B1, which is known to express recombinant proteins at the highest levels, were almost the same and the former was a little bit better. The new cell line can efficiently support the expression of foreign protein and is designated as HNU Tn FB1.