人转铁蛋白受体及其特异性抗体的制备

Preparation of the Human Transferrin Receptor and its Special Antibodies

目的:从胎盘中提取转铁蛋白受体并获得抗转铁蛋白受体的抗体。方法:人新鲜胎盘组织被破碎后,用去污剂TritonX-100裂解细胞膜,释放膜蛋白。利用膜蛋白中的转铁蛋白受体能与铁-转铁蛋白复合物特异性结合的特性对其进行亲和纯化。对纯化得到的目的蛋白,经脱盐后进行ELISA及肽质量图谱分析,证明为所需的转铁蛋白受体后,以其包被免疫管,从全合成人源噬菌体抗体库中筛选抗体。结果:从人源噬菌体抗体库中筛选到5个能够与转铁蛋白受体特异性结合的噬菌体单链抗体。结论:以人源转铁蛋白受体为抗体,可从全人源噬菌体抗体库中筛选到其特异性的抗体。

Objective： To obtain transferrin receptor（TfR） and its antibody. Methods： Human placental tissue were cut into small pieces and homogenized. Triton X-100 was added to a final detergent concentration of 2% to lysis cell. The rude extract was purified by affinity chromatography with transferrin sepharose 4B agarose and then analysized by ELISA and peptide mass fingerprint. Screening the antibody against TfR by proteins coupled to immunotubes from a full-synthesized human phage antibody library containing 10^10 different clones. Results： The results showed that the purified products were transferrin receptor and five single chain antibodies with high affinity and specificity for human transferrin receptor were selected. Conclusion： Antibodies specifically binding to nature TfR can be obtained by screening the human phage library.