中文摘要：

目的制备高效价人乳头瘤病毒（HPV）16型次要结构蛋白（L2）特异性兔血清抗体,并对其效价及其与HPV16 L2蛋白结合的特异性进行鉴定。方法将重组质粒PGEX-4T-1-HPV16 L2转化至E.coliBL21（DE3）中,经异丙基-β-D-硫代半乳糖苷（IPTG）诱导蛋白表达,SDS-PAGE和Western blot鉴定融合蛋白的抗原性;经镍螯合亲和层析柱（Ni-NTA Agarose）纯化HPV16 L2全长融合蛋白,纯化蛋白用佐剂乳化后经日本大耳白家兔背部皮下多点免疫注射,间隔2周后加强免疫,共2次,并分别于免疫前和每次免疫后1周取血,采用Western blot检测血清抗体的特异性,采用ELISA法检测血清抗体水平及变化趋势。结果家兔用HPV16 L2融合蛋白全程免疫后均产生了抗HPV16 L2抗体,ELISA法检测最高滴度达1∶256 000;Western blot检测显示,该抗体能识别HPV16 L2蛋白。结论用原核HPV16 L2全长蛋白免疫家兔,可获得高效价的血清抗体,该抗体具有与HPV L2蛋白结合的特异性。

英文摘要：

Objective To prepare a high-titer rabbit specific serum antibody against Human Papillomavirus（HPV） 16 L2 protein and identify the serum antibody titer and the binding specificity with HPV16 L2 protein.Methods The recombinant plasmid PGEX-4T-1-HPV16 L2 was transformed into E.coli BL21（DE3） and induced with IPTG.The fusion protein containing HPV16 L2 was expressed and analyzed with both SDS-PAGE and Western blot and purified with Ni-NTA Agarose.HPV16 L2 protein in Freund＇s adjuvant was prepared by the usual method.Three rabbits were immunized with the prepared antigen by subcutaneous injection at various sites on the back every two weeks,three times in all.The sera of these immunized rabbits were collected separately for preparation of rabbit serum IgG.Specific binding of the rabbit serum antibody and the HPV16 L2 protein was analyzed with Western blotting and the titer was assayed with ELISA.Results All of the immunized rabbits produced high-titer serum antibody after the second booster.The highest titer of antibody against HPV16 L2 protein was 1∶256 000.Western blotting revealed that rabbit serum antibody bound specifically to the HPV16 L2 protein.Conclusion A specific rabbit serum antibody against HPV16 L2 antigen with a high titer was successfully prepared and had binding specificity with HPV16 L2 protein.