CAJ | 학술논문

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105.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-ATT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.
목적 용α1-항이단백매(α1-antitrypsin,α1-AT)면역가토이산생다극륭항체,병대산생적항체진행유효지순화화효개감정,건립항α1-AT다극륭항체적생산공예.방법 안조상규방법면역건강가토진행항α1-AT다극륭항체적제비,용ELISA검측혈청항체효개.처사항혈청효개합격적가토,취혈、리심,획취항혈청.연합응용신산-류산안침정법화A단백친화층석법대항혈청진행순화,순화적다극륭항체경15%십이완기류산납.취병희선알응효전영분석순도후,용ELISA측정효개.결과 ELISA검측표명,가토산생적항α1-AT다극륭항체효개대우105.연합응용신산-류산안침정법화A단백친화층석법진행순화획득료순도고、활성호적항α1-ATT다극륭항체.결론 성공건립료항α1-AT다극륭항체적생산화순화공예.
Objective To prepare the polyclonal 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 polyclonai antibodies were analyzed by 15%SDS-PAGE,and antibody titers weIe measured by ELISA.Results ELISA assay showed that the titers of antisera were over 105.The polyclonal 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.