CAJ | 학술논문

目的在大肠杆菌中表达和纯化一种新型的人 IFNα 1c。方法首先建立了表达和纯化 IFNα 1c的实验室生产流程。根据干扰素的抗病毒、抗细胞增殖以及免疫调节特性，体外测定这种 IFN的生物学活性。结果 ELISA和 Western免疫印迹均证实，表达产物具有干扰素的免疫反应性。 VSV-WISH系统的抗病毒测活表明，表达的 IFNα 1c (3.2× 107) 的抗病毒活性较 IFNα 1b(1.0× 107～ 1.8× 107)略高 ;而抗 A549细胞的增殖能力则与后者相当。此外 IFNα 1c还能增强 NK细胞的杀伤活性。结论本系统成功地在大肠杆菌中表达了人 IFNα 1c。这种高比活性的干扰素可能成为临床治疗的侯选者。
목적재대장간균중표체화순화일충신형적인 IFNα 1c。방법수선건립료표체화순화 IFNα 1c적실험실생산류정。근거간우소적항병독、항세포증식이급면역조절특성，체외측정저충 IFN적생물학활성。결과 ELISA화 Western면역인적균증실，표체산물구유간우소적면역반응성。 VSV-WISH계통적항병독측활표명，표체적 IFNα 1c (3.2× 107) 적항병독활성교 IFNα 1b(1.0× 107～ 1.8× 107)략고 ;이항 A549세포적증식능력칙여후자상당。차외 IFNα 1c환능증강 NK세포적살상활성。결론본계통성공지재대장간균중표체료인 IFNα 1c。저충고비활성적간우소가능성위림상치료적후선자。
Aim To express and purify a new sub-type of interferon α , α 1c in E.coli. Methods To establish a brief protocol for expression and purification of IFN-α 1c. In vitro assay for the IFN activity is required for the measurement of antiviral, antiproliferative and immunoregulatory properties. Results Expression of IFN-α 1c was verified by its reactivity to IFN-α 1-specific neutralizing antibodies by ELISA. Western blot also detected a band with relative molecular mass(Mr) of 19 000. IFN-α 1c possessed antiviral biological activity(3.2× 107) higher than IFN-α 1b(1.0× 107～ 1.8× 107) did on VSV challenged WISH cells. IFN-α 1c also inhibited growth rate of A549 comparable to IFN-α 1b. IFN-α 1c as an immunologic effector also augmented the cytotoxic activity of NK cells. Conclusion These results demonstrate that IFN-α 1c can be successfully expressed in E.coli. This interferon with high bioactivity may be a good candidate for clinical use.