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目的 建立抗甲状腺刺激性免疫球蛋白单链抗体噬菌体抗体库,为进一步获得功能性抗体可变区基因,制备单链抗体和单链抗体-辣根过氧化物酶融合蛋白,用于甲状腺自身免疫性疾病的治疗和检测打下基础。方法 利用RT-PCR技术从分泌抗甲状腺刺激性免疫球蛋白的杂交瘤细胞中扩增出轻重链可变区基因,与噬粒表达载体p3SCMH连接,转化大肠杆菌,以甲状腺刺激性免疫球蛋白阳性患者血清IgG为抗原对表达的噬菌体抗体进行筛选。结果 经过三轮“吸附-洗脱-富集”筛选过程,洗脱下来的噬菌体呈现明显富集,收获率增加了40倍。结论 初步建立了抗甲状腺刺激性免疫球蛋白单链抗体的噬菌体抗体库。
Objective To establish anti-thyroid stimulating immunoglobulin single-chain antibody phage antibody library, in order to further obtain functional antibody variable region genes, preparation of single chain antibody and single chain antibody - horseradish peroxidase fusion protein for thyroid autoimmunity Laying the foundation for the treatment and testing of sexually transmitted diseases. Methods The heavy and light chain variable region genes were amplified from hybridoma cells secreting anti-thyroid stimulating immunoglobulin by RT-PCR and ligated with phagemid expression vector p3SCMH. The recombinant plasmid was transformed into Escherichia coli. Thyroid-stimulating immunoglobulin Patient serum IgG is an antigen that screened for expressed phage antibodies. Results After three rounds of “adsorption - elution - enrichment” screening process, the eluted phage showed obvious enrichment, the yield increased by 40 times. Conclusion The anti-thyroid stimulating immunoglobulin single-chain antibody phage antibody library was initially established.