重组人GM-CSF的理化特性及其肽图的研究

rhGM-CSF在经过盐酸胍处理,DTT还原,碘乙酰胺封闭二硫键等各种状态下,分别分析其理化特性,发现经过DTT处理后其在紫外特征光谱和RP-HPLC行为上表现出不均一性,而在SDS-PAGE电泳行为和免疫学特性方面却没有明显差异,盐酸胍对其上述指标的影响是可逆的;经过打开二硫键,封闭巯基后的肽图分析比未经处理的样品溴化氰裂解更完全,蛋白酶消化更彻底。

Physical and Chemical Characteristics and Peptide Mapping of Recombinant Human GM-CSF

The plysical and chemical characteristics of recombinant human GM - CSF (rhGM - CSF) were studied separatly. rhGM - CSF was treated by GdHCl, reduced by DTT, and the disulfide bond was blocked by idoacetamide. The results showed that the samples aren' t homogeneous in UV absorption spectrum and RP-HPLC analysis after treatment by DTT. There was no remarkable differences in the results of the analysis of SDS-PAGE electrophoresis and western blot between treated and untreated rhGM - CSF samples. The effect of GdHCl on GM-CSF was reversible in all above tests; In peptide mapping analysis, the digestion of the samples with blocked disulfide bond by CNBr and protease is more complete than that without any treatment.