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目的建立免疫毒素ScFv(anti HER2)-PE38的分离纯化工艺。方法免疫毒素ScFv(anti HER2)-PE38变性后,进行金属螯合柱层析复性和纯化,再用分子筛层析进行精纯,SDS-PAGE分析纯化产物。结果包涵体形式存在的免疫毒素ScFv(anti HER2)-PE38以8 mol/L尿素溶解效果较好;当复性梯度为稀释液从0到100%,共用时150 min,流速为2 ml/min时,复性效果较好,复性后的目的蛋白纯度可达95%以上;以凝胶柱Superdex 200进行分子筛层析效果较好,纯化产物的纯度可达98%以上。结论已建立了免疫毒素ScFv(anti HER2)-PE38的分离纯化工艺,为进一步的研究奠定了基础。
Objective To establish an isolation and purification process of anti-HER2-PE38. Methods The immunotoxin ScFv (anti HER2) -PE38 was denatured and then refolded and purified by metal chelate column chromatography. The purified product was purified by molecular sieve chromatography and analyzed by SDS-PAGE. Results The inclusion body anti-HER2-PE38 dissolved in 8 mol / L urea had a good effect. When the refolding gradient was from 0 to 100%, and the common time was 150 min, the flow rate was 2 ml / min , The refolding effect is better, the purity of the target protein after refolding can reach more than 95%; the molecular sieve chromatography with Superdex 200 gel column is better, and the purity of the purified product can reach more than 98%. Conclusion The isolation and purification of immunofluorescent anti-HER2-PE38 has been established and laid the foundation for further research.