抗人类免疫缺陷病毒gp41和丙型肝炎病毒NS3杂交瘤细胞株的建立

Establishment of hybridomas secreting monoclonal antibodies to HIVgp 41 and HCVNS 3

目的为检测血清中人类免疫缺陷病毒（ＨＩＶ）和丙型肝炎病毒（ＨＣＶ）抗原提供血清学指标。方法用纯化的重组ＨＩＶ（ｇｐ４１）和ＨＣＶ（ＮＳ３）抗原蛋白作为联合免疫原，免疫ＢＡＬＢ／ｃ小鼠，取脾细胞与Ｓｐ２／０小鼠骨髓瘤细胞融合，并采用挑单个细胞的方法进行一次克隆。结果分别获得４株和６株能稳定分泌高效价抗ＨＩＶ（ｇｐ４１）和ＨＣＶ（ＮＳ３）抗原蛋白单克隆抗体杂交瘤细胞株。初步建立了双抗体夹心法检测ＨＩＶ（ｇｐ４１）和ＨＣＶ（ＮＳ３）抗原的酶联免疫吸附试验。结论本方法是建立单抗杂交瘤细胞株的快速方法，所建杂交瘤细胞株特异性强，效价高，分泌抗体性能稳定，有推广价值。

Objective To establish the monoclonal antibodies of HIV and HCV for developing serological diagnostic method. Methods The antigens of HIV gp41 and HCV NS3 expressed from bacteria were mixed to immunize BALB/c mice. The hybridoma cell lines were obtained by fusing SP2/0 myeloma cells with spleen cells from immunized mice. Results Four hybridoma cell lines secreting specific antibodies against gp41 of HIV and six hybridoma cell lines secreting antibodies to NS3 of HCV were obtained. ELISA sandwich method for detection of viral antigens with McAbs was developed. Conclusion A quick and convenient method for obtaining McAbs was developed using mixture of different antigens for immunizition of mice. The monoclonal antibodies had high senusitivity and were stably secreted by hybridoma cell lines. This technique is worthy to be popularized.