抗氯霉素单克隆抗体杂交瘤细胞株的制备及筛选  

Preparation and screening of monoclonal antibody to chloramphenicol

作　　者：林月霞[1] 董斌[1] 袁茵[1] 段涛[1] 田素娟[1] 

机构地区：[1]广东药学院生命科学与生物制药学院,广东广州510006

出　　处：《广东药学院学报》2009年第5期526-529,共4页Academic Journal of Guangdong College of Pharmacy

基　　金：广东省科技攻关计划资助项目(2006B20501005);广州市科技计划资助项目(2006JI-C0221);广东省医学科学技术研究基金(A2006323)

摘　　要：目的获得稳定、高效分泌抗氯霉素单克隆抗体的杂交瘤细胞株。方法以CAP-HS-BSA免疫BALB/c小鼠,用细胞融合技术筛选抗氯霉素单克隆抗体杂交瘤细胞,体内诱生腹水法大量制备单抗。捕获ELISA法测定小鼠免疫球蛋白亚型,间接ELISA法测定腹水效价;用两步硫酸铵法和G蛋白亲和层析法来纯化腹水。结果成功建立了一株分泌抗CAP单克隆抗体（McAb）的杂交瘤细胞4D10。经检测,其分泌的抗体亚类为IgG1,杂交瘤染色体数目90-110条,间接ELISA检测诱生小鼠腹水的抗体效价达1∶2.56×105,纯度达95%,该细胞连续培养生物学性状稳定。结论成功制备的抗氯霉素单克隆抗体4D10效价高、纯度高,为利用该单抗研制检测氯霉素残留试剂盒提供原材料。Objective To acquire stable and secretory monoclonal antibodies against chloramphenicol hybridoma cell lines. Methods BALB/c mice were immunized with CAP-HS-BSA and hybridoma lines secreting monoclonal antibody against chloramphenicol （ CAP McAb） were screened by cell fusion, and the immunological titers of CAP McAb were determined. Capture ELISA method was used to determine isotypes of the mouse monoclonal antibody and the indirect ELISA method was performed to determine the titers of McAbs. Two-step ammonium sulfate precipitation and protein G affinity chromatography protein purification methods were used to purify ascites. Results One hybridoma cell line secreting CAP McAb 4D10 was established. The titer of purified McAb 4D10 was 1 ： 2. 56 × 10^5 , and the purity was 95%. Conclusion The anti-chloramphenicol McAb 4D10 prepared had high titer and high purity, and it could be used to develon CAP immunoassav kit for detection of CAP residues.

关 键 词：氯霉素 细胞融合 单克隆抗体 间接ELISA 

分 类 号：R392[医药卫生—免疫学]