摘要
目的筛选IL-2Rα模拟表位肽,为研制高效、特异性强的小分子肽类免疫抑制剂奠定基础。方法应用离子交换层析法从小鼠腹水中纯化抗人IL-2Rα单克隆抗体5G1,细胞ELISA方法检测其特异性。用5G1单抗筛选噬菌体环七肽库,并对噬菌体克隆进行抗原性鉴定。根据阳性克隆的DNA序列合成环肽并鉴定其生物活性。结果经5轮筛选、鉴定,获得5个与5G1有较强结合特性的噬菌体阳性克隆。DNA测序并推导氨基酸序列,得到Lys-X-{X}-Lys-Gly保守序列。依此序列合成环七肽CP,小鼠淋巴细胞增殖试验证实其有免疫抑制活性。结论环肽CP为IL-2Rα模拟表位肽,可作为IL-2Rα的拮抗剂发挥免疫抑制效应。
Objective To screen IL-2Rα mimotope that would be useful in making small peptide drugs as immunosuppressant. Methods Anti-IL-2Rα monoclonal antibody 5G1 （SGlmAb） was purified from mouse ascites by using ion exchange chromatography method and its speciality was identified by ELISA. A c7c phage display peptide library was bioanned with target molecule of 5GlmAb and the speciality of phage clones was identified. Based on the DNA sequence of positive clones, the cyclic peptide was synthesized and its biological activity was investigated. Results After 5 rounds of screening, 5 phage clones which could highly react with 5G1 were identified as the positive clones and the conservative sequence （Lys-X-{X}-Lys-Gly） was obtained. The cyclic peptide was synthesized and named CP, which had immonosupperesire activity comfinned by the lymphocyte proliferation experiment. Conclusion CP is the mimotope of IL-2Rα and it can be made as an immunosuppressant.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第1期55-57,61,共4页
Immunological Journal
