Comparative Proteomics Analysis Of Human Blood Components And Derivates

With the accomplishment of Human Genome Project, the research of life science has been entering Post-genome era. Now proteomics research has become one of the hot fields of 21 century life science. In this paper, we constructed plasma proteomic research platform based on two-dimensional electrophoresis (2-DE) and performed series of comparative proteomic research for human blood components and their derivates containing plasma, serum, blood cell, and human albumin etc.Aiming at normal human plasma, we established and optimized 2-DE method and information processing method for plasma proteomics. These include: 1) established acidic silver stains as basic method to visualize gels through comparing the sensitivity of diamine silver stains and acidic silver stains, whose sensitivity is as high as 2ng and whose background is lower than diamine silver stains; 2) improved the clarity and stability of 2-DE gels through adjusting and optimizing electrophoresis parameters of Isoelectric Focusing; 3) appraised removal effect of Albumin and IgG removal Kit. We found that this kit has significant effect to remove albumin and globin in plasma. But it removes low abundance proteins nonspecifically in the same time; 4) through comparing the separation effect of different IPG strips with different pH, established that the separation effect of narrow pH (4-7) IPG strips is obviously better than that of wide pH (3-10) IPG strips; 5) established the method to identify differential spots by combing auto detection function of software and intensity analysis.Normal human plasma, domestic human albumin products and domestic recombinant human serum albumin were separated respectively by SDS-PAGE and 2-DE and detected by the colloidal Coomassie brilliant blue and silver stains. With the help of Labworks45 software, the purity of human serum albumin was compared. The results show that: 1) the purity of human serum albumin detected by SDS-PAGE and 2-DE is lower than that detected by current method. 2) recombinant human serum albumin contains much protein from host cells. These results indicate that SDS-PAGE and 2-DE may improve sensitivity for analysis of blood products.The proteomic difference of plasma between normal human (control group) and SARS convalescent patients (experimental group) which has different titer of neutralizing antibody was compared by 2-DE method. The results show that: 1)...