中文摘要：旨在制备抗磺胺二甲氧嘧啶 ( SDM) 驼源单域重链 ( VHH) 抗体，用于检测动物源性食品中 SDM 的残留。采用重氮化法，SDM 分别与牛 血清白蛋白 ( BSA) 和鸡卵清蛋白 ( OVA) 偶联，合成人工免疫原 ( SDM－BSA) 和包被抗原 ( SDM－OVA) 。用 SDM－BSA 免疫骆驼，在第 5 次 免疫后 1 周采集骆驼外周血液，分离外周血淋巴细胞，提取 ＲNA，ＲT－PCＲ 扩增 VHH 基因，将 VHH 基因插入 pCANTAB－5E 噬菌粒载体，构建 双峰驼单域重链抗体库。经本实验室前期测定其库容为 1. 08×105 CFU，阳性率为 96. 6%。利用噬菌体展示技术经过 4 轮生物淘选，筛选获得特异 性表达抗 SDM 抗体的重组噬菌体。结果: 构建了 SDM 的驼源 VHH 抗体库，经生物淘选及 phage ELISA 检测获得了特异性的重组噬菌体。通过 4 轮生物淘选，获得了能够与 SDM－OVA 抗原有明显结合力且能够特异性表达抗 SDM 抗体的重组噬菌体，为后期检测 SDM 在动物源性食品残留奠 定了良好的基础。
外文摘要：To prepare a camel－derived variable domain of heavy－chain antibody ( VHH) against sulfadimethoxine ( SDM) for the detection of SDM residues in animal foods． Diazotization was used to couple SDM with bovine serum albumin ( BSA) and chicken ovalbumin ( OVA) to synthesize artificial immunogen SDM－BSA and coated antigen SDM－OVA． Experimental camels were immunized with SDM－BSA，and peripheral blood samples were collected from the camels one week after the fifth immunization，peripheral blood lymphocytes were isolated， ＲNA was extracted，and the VHH gene was amplified by ＲT－PCＲ． The VHH gene was inserted into the pCANTAB－5E phage vector to construct a bactrian camel heavy chain antibody library with a laboratory pre－determined storage capacity of 1. 08×105 CFU and the positive rate was 96. 6%． After four rounds of biological panning using the phage display technology，specific recombinant phages were screened． As a result，a camel－derived anti－SDM VHH antibody library was constructed specific recombinant phages were obtained by bio－panning and phage ELISA detection． The recombinant phage that could bind to the SDM－OVA antigen and specifically express anti SDM antibody was obtained through four rounds of biological panning，which laid a good foundation for the subsequent experiments．
中文关键词：磺胺二甲氧嘧啶 ( SDM) ；单域重链抗体 ( VHH) ；噬菌体展示技术
外文关键词：sulfadimethoxine ( SDM) ；variable domain of heavy chain of heavy－chain antibody ( VHH)；phage display technology
作者：贺东阳；张齐；张星星；钟发刚；张小莺；王金泉
作者单位：新疆农业大学；新疆农垦科学院；Department of Biomedical Sciences
期刊名称：畜牧与兽医
期刊影响因子：0.678
出版年份：2021
出版刊次：1
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