摘要
目的用α1-抗胰蛋白酶（α1-antitrypsin，α1-AT）免疫家兔以产生多克隆抗体，并对产生的抗体进行有效地纯化和效价鉴定，建立抗α1-AT多克隆抗体的生产工艺。方法按照常规方法免疫健康家兔进行抗α1-AT多克隆抗体的制备，用ELISA检测血清抗体效价。处死抗血清效价合格的家兔，取血、离心，获取抗血清。联合应用辛酸-硫酸铵沉淀法和A蛋白亲和层析法对抗血清进行纯化，纯化的多克隆抗体经15％十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析纯度后，用ELISA测定效价。结果ELISA检测表明，家兔产生的抗α1-AT多克隆抗体效价大于10^5。联合应用辛酸．硫酸铵沉淀法和A蛋白亲和层析法进行纯化获得了纯度高、活性好的抗α1-AT多克隆抗体。结论成功建立了抗α1-AT多克隆抗体的生产和纯化工艺。
Objective To prepare the polyelonal antibody against α1 -antitrypsin （ α1-AT） by immunizing rabbits with α1 -AT, and to purify and characterize the antibodies produced from rabbits in order to establish the production process of the polyclonal antibody against α1-AT. Methods The healthy rabbits were routinely immunized with α1-AT to prepare polyclonal antibody against α1-AT. The serum antibody titers were determined by ELISA. The rabbits with qualified antibody titers were killed to collect blood. The antisera, obtained by centrifugalizing the whole blood, were purified by caprylic acid-ammonium sulfate precipitation and protein A affinity chromatography. The purified polyclonal antibodies were analyzed by 15% SDS-PAGE, and antibody titers were measured by ELISA. Results ELISA assay showed that the titers of antisera were over 105. The polyelonal antibodies, purified by caprylic acid-ammonium sulfate precipitation and protein A affinity chromatography, had good purity and immunological activity. Conclusion The production and purification processes of polyclonal antibody against α1-AT are successfully established.
出处
《国际生物制品学杂志》
CAS
2009年第3期122-124,共3页
International Journal of Biologicals
关键词
Α1抗胰蛋白酶
多克隆抗体
制备
纯化
鉴定
Alpha 1-antitrypsin
Polyclonal antibody
Preparation
Purification
Characterization
