原核和真核细胞表达HBeAg的应用研究

目的：将原核细胞和真核细胞分别克隆表达生产的HBeAg，经适当纯化后进行检测分析，并在乙型肝炎抗HBe检测中进行应用和比较。方法：分别用大肠杆菌和家蚕细胞表达生产HBeAg，并用Saphacryl S-200柱层析进行纯化；紫外分光光度法测定表达产物的蛋白含量；EIA法测定HBeAg和HBcAg效价及评估HBeAg的应用效果。结果：原核细胞HBeAg：比活性为10000／mg,HBeAg/HBcAg＝50，用于抗HBe的检测时特异性为96％，灵敏度符合国家卫生部panel要求，真核细胞HBeAg：比活性为160000／mg,HBeAg/HBcAg＝5000，用于抗HBe的检测时特异性为100％，灵敏度高于国家卫生部panel要求的1－2个滴度。结论：真核细胞表达的HBeAg比活性高HBcAg含量低，在抗HBe检测时的应用效果优于原核细胞。

Application of HBeAg produced in prokaryotic cells and eukaryotic cells

Objectives: To express HBeAg in prokaryotic and eukaryotic cells and compare the two types of HBeAg in the anti HBeAg testing. Methods: HBeAg was expressed both in E.coli cells and in silk worm cells, purified by Sephacryl S 200.HBeAg protein concentration and antigenic titer were determined respectively by ultraviolet spectroscopy and EIA. Results: HBeAg produced by E.coli cells: Activation ratio was 10 000/mg, HBeAg/HBcAg = 50; The specificity in testing anti HbeAg was 96%;HBeAg produced by silk worm cells: Activation ratio was 160 000/mg, HBeAg/HBcAg = 5 000, The specificity in testing anti HbeAg was 100%. Conclusions: HBeAg produced by eukaryotic cells contained much lower proportion of HbcAg and higher activation ratio, which therefore bring about a possibility to improve the quality of the kit for testing Anti HBe.