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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。
OBJECTIVE: To construct and prepare a novel sequence of monoclonal antibody MAB1 which blocks the function of p40 of interleukin-12 (IL-12) by antibody engineering and to identify its biological activity, so as to prepare a novel monoclonal antibody MAB1 which can be used in the treatment of various autoimmune diseases IL-12 pathway blocker. Methods According to the existing IL-12 p40 protein sequence, a novel monoclonal antibody MAB1 was designed, constructed and prepared. The affinity of antigen-binding MAB1 was measured by indirect ELISA and its biological activity was identified by cytology. Results The binding affinity of antibody MAB1 to p40 antigen was 0.0399 nmol / L, and the level of IL-12-induced human skin lymphocyte-associated antigen (CLA) was significantly inhibited. Its antigen binding ability and the ability to inhibit the biological activity of IL-12 are comparable to or better than the US Johnson & Johnson antibody Stelara (ustekinumab). Conclusion The newly constructed monoclonal antibody MAB1 acts as a high-affinity blocker of the IL-12 pathway and thus becomes a clinical candidate drug for the treatment of new autoimmune diseases such as plaque psoriasis.