摘要
确定重组HBV表面抗原和HCV核心抗原对奶牛的最适免疫剂量、免疫次数及间隔时间,观察牛奶中抗HBV与抗HCV的效价与消长规律。表达HBV表面抗原和HCV核心抗原的重组菌经IPTG诱导后,目的蛋白以包涵体形式存在。放大发酵工艺,菌体发酵密度达40g/L,目的蛋白表达量为26%～30%。将包涵体变性、复性后,测定蛋白含量,并用SDS-PAGE鉴定。采用不同的剂量和不同的抗原处理方法免疫奶山羊和奶牛,检测HBV抗体、HCV抗体、干扰素与白介素等细胞因子。免疫奶山羊羊奶中的HBV抗体效价最高为1∶80,HCV抗体效价最高为1∶20;6个月后,HBV抗体效价无明显下降,HCV抗体效价下降明显。对奶牛5个批次的免疫结果显示,免疫次数应多于3次,免疫剂量以每头牛300μg为宜,间隔时间以1个月为宜。免疫牛奶中HBV抗体的阳性率约为66.0%,抗体效价最高为1∶160;HCV抗体阳性率约为17.0%,维持时间较短;孕牛比旺奶牛产生抗体的效价高。免疫牛奶中检测到了干扰素和白介素等细胞免疫活性因子。
The recombinant E.coli expressing HBsAg and HCV core antigen was induced by IPTG. The fermented density of the recombinant E.coli was 40 g/L, and the expressed protein existed in inclusion bodies accounted for 26%-30% in the total protein. The inclusion bodies were identified by SDS-PAGE after denaturation and renaturation. The milk goat and the milch cow were immunized with different doses and artifactitious antigen. The anti-HBs, anti-HCV, IFN-α and IL-1 were examined by ELISA. The anti-HBs dilution was 1：80 and the anti-HCV dilution was 1：20 in goat milk. The anti-HBs remained and the anti-HCV descended distinctly after 6 months in goat milk. Then the anti-HBs positive rate was 66.0%, the anti-HCV positive rate was 17.0% and the anti-HBs dilution was 1：160 in milk. The best immune project was as follow： over 3 times, 300μg each milch cow when every injection, 30 d internal. The antibody dilution in pregnancy cow milk was higher than that in general milch cow. The anti-HBs remained longer than the anti-HCV in milk. IFN-α and IL-1 were detected.
出处
《生物技术通讯》
CAS
2005年第6期632-634,共3页
Letters in Biotechnology
基金
国家农业科技成果转化重点项目(02EFN216100573)
作者简介
尹文（1969-），男，副教授，博士，（E-mail）yinwen@fmmu．edu．cn
