EB病毒膜蛋白家兔免疫血清的制备及其特性研究

目的：制备高效价EB病毒膜蛋白抗体,并对其效价及其与EBV结合的特异性进行鉴定。方法：将B95-8细胞培养至3×10^5/mL时,超声破碎,纯化获得EB病毒膜蛋白抗原;EB病毒膜蛋白抗原经弗氏佐剂乳化,在家兔背部皮下多点免疫注射,间隔2周免疫,共3次,并分别于免疫前和每次免疫后1周取兔血清,检测兔免疫血清抗体IgG;进一步采用Western blot检测制备的血清抗体与EBV结合的特异性,同时采用ELISA法检测兔血清抗体水平及各周抗体滴度的变化趋势。结果：所有家兔在全程免疫后都产生了高效价膜蛋白的特异性抗体,最高滴度达1：6400。结论：用B95-8细胞制备的EB病毒膜蛋白抗原免疫家兔,可成功制备高效价的特异性血清抗体,为EB病毒膜蛋白抗原的免疫学检测奠定了基础。

Preparation and research on immunological characteristics of rabbit serum antibody to EBV membrane protein

Objective： To prepare bigh-titer rabbit serum antibody against EBV membrane protein. Methods： B95-8（EBV） cells were cultivated and proliferated in cell culture system with RPMI-1640 culture medium （with 10% FBS and Pen/Strep）. Thereafter, the purified B95-8（EBV） was harvested using supercentrifuge method. Prepare EBV membrane protein containing Freund adduvant by routine method. Four rabbits were immunized with the prepared antigen by subcutaneous injection at various sites of skin on back for every two weeks,three times in all. The sera of these immunized rabbits were collected separately for preparation of the rabbit serum IgG. Specific binding of the rabbit serum antibody and the EBV membrane protein were analyzed by Western blot and the titer was assayed by ELISA. Results： All of the immunized rabbits produced high-titer serum antibody after the second booster. The highest titer of antibody against EBV membrane protein was 1：6 400. The result of Western blot showed the rabbit serum antibody could bind to the EBV membrane protein specifically. Conclusion： The specific rabbit serum antibody against EBV membrane antigen,with high titer,is successfully prepared. It laid a foundation of development of immunology method for detection of EBV membrane antigen.