抗氯霉素单克隆抗体的制备与纯化
Preparation and purification of monoclonal antibodies against chloramphenicol.
作者信息
Yi Yu, Wang Zhuhuan, Li Min, Zhu Keyin, Ying Guoqing
机构信息
College of Pharmaceutical Science, Zhejiang University of Technology, Chaowang road 18, Hangzhou, 310014, China.
出版信息
Cytotechnology. 2012 Mar;64(2):157-63. doi: 10.1007/s10616-011-9401-6. Epub 2011 Dec 11.
Monoclonal antibodies (McAbs) against chloramphenicol (CAP) were produced to detect CAP residues, which could be toxic and possesses a potential threat to human health. The CAP-BSA conjugate was obtained by bovine serum albumin (BSA) coupled with CAP, and used to immunize the mice. The splenocytes from the immunized mice were fused with mouse myeloma cells SP2/0 to form hybridoma, which may secrete McAbs against CAP. Hybridomas 1D(1) and 3G(12) secreting McAbs against CAP were obtained by screening. Ascites containing McAbs were prepared by injecting 1 x 10(6) cells of hybridoma 1D(1) and 3G(12) into the abdomen of mice. Protein A affinity chromatography was used to purify McAbs against CAP in a single chromatographic step with recovery yield above 80% and purity above 95% and full recovery of antibody activity. Experiments showed that McAb 3G(12) was highly specific for CAP and had no cross-reactivity with analogues which have a structure similar to CAP. The IC(50) value was 50.8 ng/mL.
制备了抗氯霉素（CAP）的单克隆抗体（McAbs）以检测CAP残留，CAP残留可能有毒并对人类健康构成潜在威胁。CAP - 牛血清白蛋白（BSA）偶联物通过牛血清白蛋白（BSA）与CAP偶联获得，并用于免疫小鼠。将免疫小鼠的脾细胞与小鼠骨髓瘤细胞SP2/0融合以形成杂交瘤，其可能分泌抗CAP的McAbs。通过筛选获得了分泌抗CAP的McAbs的杂交瘤1D(1)和3G(12)。通过将杂交瘤1D(1)和3G(12)的1×10(6)个细胞注射到小鼠腹腔中来制备含有McAbs的腹水。采用蛋白A亲和色谱法在单个色谱步骤中纯化抗CAP的McAbs，回收率高于80%，纯度高于95%，抗体活性完全恢复。实验表明，McAb 3G(12)对CAP具有高度特异性，与结构类似于CAP的类似物无交叉反应。IC(50)值为50.8 ng/mL。
Zotero 插件