可捕获HBV的preS1单克隆抗体的制备及性质初探

Isolation and Characteration of the Anti-native HBV preS1 Monoclonal Antibodies

用天然乙型肝炎病毒（HBV）颗粒（Dane颗粒和管状颗粒）和HBV前S1（preS1）区基因工程重组蛋白GST-preS1联合免疫Balb/c小鼠，经杂交瘤技术制备了7株单克隆抗体（mAb），采用ELISA、免疫捕获PCR等方式鉴定其有关特性.各株mAb分别为IgG1和IgG2a，轻链均为κ型.腹水mAb的滴度为1×10^7-1×10^9.用间接法ELISA显示，所有7株preS1 mAb均可以识别HBV天然抗原，其中mAb 4D11与HBV天然抗原的结合能力最强.通过竞争抑制法ELISA检测推断preS1（21-47）上至少含有两个B细胞表位，mAb 4D11、1G5、7B6和7H11具有共同的B细胞抗原识别位点；mAb 3H5、6F1和2A6识别另外一个位点.本研究获得的可与HBV天然抗原结合的preS1单抗为HBV preS1检测试剂的研制提供了重要工具，对HBV preS1具有中和免疫活性抗原的设计及相关研究有重要帮助.

The Balb/c mice were immunized with native HBV antigen （Dane particles and tubular particles）and purified recombinant protein GST-preS1. Seven mAbs anti-HBV preS1 mAbs were prepared by hybridoma technique, and the relative properties of the mAbs were identified by ELISA and immu-capture PCR. Of these mAbs, mAb 4D11 and 7H11 were IgG1, the others were IgG2a. The isotype of mAb 7B6 light chains were lamda （λ）, the others were kappa （κ）. All of these mAbs could recognize native HBV antigens （Dane particles and tubular particles）, luther study shows that they could recognize two different B cell epitopes of preS1 （21- 47） ,one recognized by mAb 4D11, 1G5,7B6 and 7H11, the other is recognized by mAb 3H5,6F1 and 2A6. The mAbs＇ abilities of recognizing the native HBV antigen show use of developing regent to test HBV preS1 ,and assisting in the design of neutralizing antigen.