Huang Yi, Hu Lei, Yang Yan-Qun, Hu Xue-Ping, Zhen Yong-Su, Liu Meng-Yuan
Center for Infection and Immunity Research, Life Sciences College of Hubei University, Wuhan 430062, China.
Yao Xue Xue Bao. 2012 Sep;47(9):1210-8.
In order to increase the plasma half-life and tissue specificity of IL-1 receptor antagonist, a recombinant fusion protein IL-1Ra-HSA, linked by a rigid peptide linker PAPAP, was engineered and expressed by the Pichia pastoris host cells. The fusion protein was secreted to the host cells culture, identified by Western blot, and purified by affinity chromatography. This was followed by a further examination of its bioactivity and pharmacokinetics. Our results demonstrated that the fusion protein retained the antagonist activity of IL-1Ra, capable of binding specifically to the IL-1 receptor on human melanoma A375.S2 cells, and inhibits the cytolytic effect of IL-1beta to A375.S2 cells. Albumin fusion dramatically extended the half-life of IL-1Ra and resulted in a specific accumulation of IL-1Ra in the arthritic paws and a lower distribution of IL-1Ra in other organs such as liver, kidney, spleen and lung in mice with collagen-induced arthritis. The findings reported herein indicate that the fusion protein is likely to have greater clinical applications in areas such as the treatment of rheumatoid arthritis.
为了提高白细胞介素-1受体拮抗剂(IL-1Ra)的血浆半衰期和组织特异性,通过刚性肽接头PAPAP连接构建了重组融合蛋白IL-1Ra-HSA,并由毕赤酵母宿主细胞表达。融合蛋白分泌到宿主细胞培养物中,通过蛋白质免疫印迹法进行鉴定,然后通过亲和层析法进行纯化。随后对其生物活性和药代动力学进行了进一步研究。我们的结果表明,该融合蛋白保留了IL-1Ra的拮抗剂活性,能够与人黑色素瘤A375.S2细胞上的IL-1受体特异性结合,并抑制IL-1β对A375.S2细胞的细胞溶解作用。白蛋白融合显著延长了IL-1Ra的半衰期,并导致IL-1Ra在胶原诱导性关节炎小鼠的关节炎爪中特异性蓄积,而在肝脏、肾脏、脾脏和肺等其他器官中的分布较低。本文报道的研究结果表明,该融合蛋白在类风湿性关节炎治疗等领域可能具有更大的临床应用价值。