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目的建立稳定表达含有pre-S2的HBsAg的细胞系。方法构建在真核细胞表达pre-S2-HB- sAg的重组质粒PCI-HBs2。用此质粒转染HepG2细胞,经克隆化培养以及G418筛选,建立稳定表达HB2Ag的细胞系。对其分泌动态、细胞纯度、遗传稳定性等生物学特性做了鉴定。同时对该细胞实验室大量培养的条件进行了优化。结果成功构建表达pre-S2-HBsAg的重组质粒。转染细胞后筛选到一个稳定表达pre-S2-HBsAg的细胞系3E3。ELISA检测表明该细胞系分泌的HBsAg水平高,生物学性状研究结果表明稳定表达HBsAg的3E3细胞纯度好,遗传性状稳定。结论成功建立了稳定表达pre-S2-HBsAg的细胞系。该细胞系遗传性质稳定,表达蛋白产量高。此细胞系可用于pre-S2-HBsAg的制备及纯化。
Objective To establish a cell line stably expressing HBsAg containing pre-S2. Methods The recombinant plasmid PCI-HBs2 expressing pre-S2-HB-sAg in eukaryotic cells was constructed. HepG2 cells were transfected with this plasmid, cloned and selected by G418 to establish a cell line stably expressing HB2Ag. Its secretion dynamics, cell purity, genetic stability and other biological characteristics were identified. At the same time, the cell culture laboratory conditions were optimized. Results The recombinant plasmid expressing pre-S2-HBsAg was successfully constructed. The transfected cells were screened to a cell line 3E3 stably expressing pre-S2-HBsAg. The result of ELISA showed that the cell line secreted high level of HBsAg. The biological characterization results showed that the 3E3 cells stably expressing HBsAg had good purity and stable genetic traits. Conclusion The cell line stably expressing pre-S2-HBsAg was successfully established. The cell line is characterized by its stable genetic profile and high protein yield. This cell line can be used for the preparation and purification of pre-S2-HBsAg.