用包涵体复性的方法制备重组人白细胞介素-1受体拮抗剂

Using Process of Refolding the Protein to Obtain Recombinant Human Interleukin-1 Receptor Antagonist

通过大肠杆菌将IL-1ra表达成包涵体,将包涵体用8M尿素溶解后,稀释4倍后直接用离子交换柱层析进行复性和纯化,复性后得到的IL-1ra纯度大于95%,生物活性大于1×105IU/mg,内毒素含量也比较低。Western-Blotting印迹也表明重组蛋白具有IL-1ra的抗原活性。

Recombinant human interleukin-1 receptor antagonist was expressed in E. coli as an insoluble inclusion body. The inclusion body was dissolved in the 8 M urea and then the solution was diluted untill the concentration of urea became 2 M. By ion exchange chromatography the protein in the solution of 2 M urea was refolded and purified. At last the purity of product is more than 95% and its bioactivity is more than 1× 10^5 IU/mg while it has little endotoxin. Western-Blotting also indicates that recombinant protein can react with antibodies against anti-hIL-1ra.