CAJ | 학술논문

目的：设计并利用抗体工程方法构建并制备全新序列的阻断白介素12（IL-12）p40功能的单克隆抗体 MAB1，并对其生物学活性进行鉴定，以制备可应用于治疗多种自身免疫病的 IL-12通路的阻断剂。方法根据已有的 IL-12 p40蛋白序列，设计、构建并制备了全新的单克隆抗体 MAB1，利用间接 ELISA 测定其与抗原结合的亲和力，并用细胞学方法鉴定它的生物学活性。结果抗体 MAB1与 p40抗原的结合亲和力为0.0399 nmol/L，并能明显抑制 IL-12诱导的人皮肤淋巴细胞相关抗原（CLA）水平上调。其抗原结合能力以及抑制IL-12生物学活性能力与美国强生公司抗体药 Stelara （ustekinumab）相当或更优。结论全新构建的单克隆抗体 MAB1可作为 IL-12通路的高亲和力阻断剂，从而成为自身免疫疾病（例如斑块型银屑病）治疗用新药的临床候选药物。
목적：설계병이용항체공정방법구건병제비전신서렬적조단백개소12（IL-12）p40공능적단극륭항체 MAB1，병대기생물학활성진행감정，이제비가응용우치료다충자신면역병적 IL-12통로적조단제。방법근거이유적 IL-12 p40단백서렬，설계、구건병제비료전신적단극륭항체 MAB1，이용간접 ELISA 측정기여항원결합적친화력，병용세포학방법감정타적생물학활성。결과항체 MAB1여 p40항원적결합친화력위0.0399 nmol/L，병능명현억제 IL-12유도적인피부림파세포상관항원（CLA）수평상조。기항원결합능력이급억제IL-12생물학활성능력여미국강생공사항체약 Stelara （ustekinumab）상당혹경우。결론전신구건적단극륭항체 MAB1가작위 IL-12통로적고친화력조단제，종이성위자신면역질병（례여반괴형은설병）치료용신약적림상후선약물。
Objective To design and produce a new sequence antibody MAB1 that binds to the p40 subunit of IL12 and blocks IL12 function, for treating autoimmune diseases mediated by IL-12 activity. Methods A monoclonal antibody MAB1 which specifically bond to IL-12 p40 was designed and constructed based on the crystal structure of IL-12 p40. The affinity of MAB1 directed p40 was assessed by indirect ELISA and inhibition of IL-12 induced cutaneous lymphocyte associated antigen (CLA) up-regulation was assessed by FACS analysis. Results In the present study, monoclonal antibody MAB1 was successfully generated. It possessed high binding affinity to p40 and potent inhibitory effect of IL-12 induced CLA up-regulation. These activities were comparible or equivalent to the properties of ustekinumab. Conclusion Monoclonal antibody MAB1 is a high affinity IL-12 antagonist and has potential to be developed for autoimmune diseases such as plaque psoriasis.