构建肿瘤坏死因子-α免疫微球的实验研究

Building Immune Microsphere against Tumor Necrosis Factor-alpha(TNF-α)

前瞻性构建肿瘤坏死因子-α免疫微球,以期为将来进一步研究能应用于特异性清除TNF-α的血液净化方法,奠定实验基础。采用以PSM为载体,依次包被PLL及rHTNF-αM cA b的方法而制备的免疫微球,分别以异硫氰酸荧光素标记,应用倒置显微镜及荧光显微镜观察、分光光度计测定等方法,探讨吸附微球的包被条件。结果显示,20℃、pH 9.5、60 m in三者为PLL包被PSM的最佳条件。包被后的洗脱液检测结果显示PLL含量无明显变化,说明PLL在PSM表面包被较为牢固。在相同温度及包被时间内,应用浓度为0.2%的戊二醛溶液进行的rHTNF-αM cA b对PLL的包被结合牢固。实验表明,所构建的肿瘤坏死因子-α免疫微球,达到了预期的目的,可以作为一种新颖的免疫吸附材料。该方法简单,价格便宜,为相关实验研究提供了一种崭新方法。

We have constructed the immune microsphere against tumor necrosis factor-alpha （TNF-α） prospectively, hoping to establish the experiment groundwork in more researches which could be used in specific elimination of the TNF-α by blood purification method for the future. The recombinant human tumor necrosis factor-alpha monoclonal antibody （rHTNF-α McAb） was wrapped on the polystyrene microsphere （PSM） carrier connecting poly-L-lysine （PLL） beforehand. They were earmarked by the fluorescein isothiocyanate （FITC） respectively. The packing conditions were examined using the inversted and fluorescence microscopes and the spectrophotometer. The results showed that the best conditions for wrapping were 20 ℃, prig. 5 and 60 minutes. The PLL content was not changed in the washing fluid after coating, which indicated the wrapping was quite firm. At the same temperature and same coating time, the rHTNF-α McAb coated on the PLL was obviously substantial when the concentration of glutaraldehyde solution was 0. 2%. The findings demonstrated that the built immune microsphers can be used as a novel adsorption material. This method is simple and economic, and it offers a new approach to the related studies.