Santos Nayara G L, Rocca Mayra P, Pereira Carlos A, Ventini Daniella C, Puglia Ana Lia P, Jorge Soraia A C, Lemos Marcos A N, Astray Renato M
Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil.
Cytotechnology. 2016 Dec;68(6):2605-2611. doi: 10.1007/s10616-016-9984-z. Epub 2016 May 23.
Recombinant Drosophila S2 cells have been used for the expression of many proteins of medical interest. However, membrane-attached glycoproteins, which commonly exhibit lower expression levels compared to soluble proteins, may require special procedures in order to attain high levels of expression. In this study, two S2 cell population enrichment methods (antibiotic and immunomagnetic selection) were evaluated for their ability to enhance expression of the membrane-anchored rabies virus glycoprotein (RVGP). Quantification of RVGP production and determination of its cDNA copy number in transformed cells showed that both enrichment methods increased RVGP expression without significantly affecting its gene copy number. More interestingly, RVGP mRNA levels measured after cycloheximide treatment were poorly correlated with glycoprotein levels. Both enrichment methods enhanced expression of RVGP by recombinant S2 cells, with the highest level of expression achieved using immunomagnetic selection.
重组果蝇S2细胞已被用于表达许多具有医学意义的蛋白质。然而，与可溶性蛋白质相比，膜附着糖蛋白的表达水平通常较低，可能需要特殊程序才能实现高水平表达。在本研究中，评估了两种S2细胞群体富集方法（抗生素和免疫磁珠筛选）增强膜锚定狂犬病病毒糖蛋白（RVGP）表达的能力。对转化细胞中RVGP产量的定量及其cDNA拷贝数的测定表明，两种富集方法均提高了RVGP的表达，而未显著影响其基因拷贝数。更有趣的是，环己酰亚胺处理后测得的RVGP mRNA水平与糖蛋白水平相关性较差。两种富集方法均增强了重组S2细胞对RVGP的表达，其中免疫磁珠筛选实现了最高水平的表达。
