Preparation and purification of monoclonal antibodies against chloramphenicol
Preparation and purification of monoclonal antibodies against chloramphenicol
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DOI：
10.1007/s10616-011-9401-6
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发表时间：
2012-03
期刊：
影响因子：
2.2
通讯作者：
Y. Yi;Zhu-ming Wang;Min Li;Keyin Zhu;Guoqing Ying
中科院分区：
文献类型：
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作者：
Y. Yi;Zhu-ming Wang;Min Li;Keyin Zhu;Guoqing Ying
氯霉素（CAP）是一种对人体健康具有潜在威胁的有毒物质，本研究制备了抗CAP的单克隆抗体（McAb）用于检测氯霉素残留。将牛血清白蛋白（BSA）与CAP偶联，制备CAP-BSA偶联物，并免疫小鼠。免疫小鼠的脾细胞与小鼠骨髓瘤细胞SP2/0融合，获得分泌抗CAP单克隆抗体的杂交瘤细胞株。经筛选获得分泌抗CAP单克隆抗体的杂交瘤细胞株1D 12和3G 12。将杂交瘤细胞1D 11和3G 12分别接种于小鼠腹腔，制备腹水。用Protein A亲和层析法一步纯化抗CAP单克隆抗体，回收率在80%以上，纯度在95%以上，抗体活性完全恢复。实验表明，3G 12单抗对CAP具有高度特异性，与CAP类似物无交叉反应。IC 50值为50.8 ng/mL。
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 1D1and 3G12secreting McAbs against CAP were obtained by screening. Ascites containing McAbs were prepared by injecting 1 x 106cells of hybridoma 1D1and 3G12into 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 3G12was highly specific for CAP and had no cross-reactivity with analogues which have a structure similar to CAP. The IC50value was 50.8 ng/mL.