中文摘要：

目的：对人未做处理的血清以及去除白蛋白和免疫球蛋白G（IgG）血清的蛋白质组学方法进行比较和优化。方法：应用双向电泳（2-DE）方法分离了未做处理的以及去除白蛋白和免疫球蛋白G（IgG）的血清，比较优化了高温变性、水化液成份组成及泡胀方式等影响血清2-DE分离效果的因素，并用质谱分析鉴定未做处理和已处理血清的2-DE谱图中部分差异蛋白点。结果：得到了分辨率和重复性较好的2-DE谱图，未做处理的血清、去除白蛋白及IgG血清的平均蛋白质点分别为（482±18）个和（523±29）个，质谱分析了9个差异蛋白点，鉴定为8种蛋白质，其中7种为功能蛋白质。仅出现在未做处理血清中的蛋白有4种，分别是维生素A结合蛋白、可溶性尿激酶血纤维蛋白溶酶原激活剂受体、蛋白激酶1抗原、血清白蛋白。4种蛋白仅出现在去除白蛋白和IgG的血清中，分别是NADH脱氢酶辅酶β亚基、肌动蛋白结合蛋白M1、T细胞活性受体β链、血小板生长因子C。结论：去除高丰度蛋白可增加一些低丰度蛋白质的检出，但非特异性吸附会导致部分功能蛋白质的丢失。

英文摘要：

Objective： To compare and optimize serum proteomics of neat human serum and removal of albumin and immunoglobulin G （IgG） serum. Methods： Two dimensional gel electrophoresis （2-DE） was utilized to compare protein expression profiles of neat human serum and removal of albumin and immunoglobulin G （IgG） serum. And various conditions of serum proteomic 2-DE, such as high temperature denaturalization, rehydration buffer and rehydration model, were adjusted and optmized. PDOuest image analysis was utilized to compare protein expression profiles of neat human serum and removal of albumin and immunoglobulin G （IgG） serum. Parts of different proteins were identified with mass spectrometry and database searches. Results： Through modified the conditions of high temperature denaturalization, rehydration buffer and rehydration model, well-resolved and reproducible 2-DE profiles were obtained. And the average spots in the gels of pH=4-7 IPG strips for neat serum and removal of albumin and IgG serum were （482±18） and （523±29）, respectively. Nine protein spots were subjected to mass spectrometry analysis and eight proteins were identified by PMF, seven proteins have biological functions. The results showed that Plasma rednol-binding protein, soluble urokinase plasminogen activator receptor, APK1 antigen and ALB protein were only expressed in neat human serum, and NADH dehydrogenase 1 beta subcomplex, acdn related protein M1, T-cell receptor active beta-chain V-D-J-beta-1.2-C-beta-1 and platelet-derived growth factor C precursor were only expressed in removal of albumin and immunoglobulin G （IgG） serum. Conclusion： Treatment of serum samples with affinity-blue gel and protein A before 2-DE analysis can be used to remove high abundance proteins in order to increase the detection sensitivity of proteins present in low abundance, but parts of low abundance proteins may be lost for unspecific binding in the process.