固相配体文库降低血清高低丰度蛋白浓度差异  

Decrease of serum protein concentration difference by solid-phase ligand libraries

作　　者：齐义军[1] 牛保华[1,2] 王攀[1] 侯艳芳[1] 晁玮霞[3] 马远方[1] DG Ward PJ Johnson 

机构地区：[1]河南大学医学院细胞与分子免疫学重点实验室 [2]河南大学环境医学研究所 [3]河南大学病理生理学教研室,河南开封475004 [4]伯明翰大学癌症研究所

出　　处：《解剖学报》2011年第6期770-774,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30700366;81072039)

摘　　要：目的探讨固相配体文库技术富集血清低丰度蛋白及提高质谱鉴定低丰度蛋白敏感性。方法3组混合血清标本采用蛋白富集珠(ProteoMiner)处理,一维或二维电泳图谱分析比较处理前后血清图谱中的差异条带或蛋白质斑点,并于处理后血清的二维电泳图谱中显著增加的蛋白质斑点进行高效液相色谱法电喷雾串联质谱鉴定。结果一维及二维电泳图谱表明,固相配体文库技术能显示更多的蛋白条带或蛋白质斑点;显著富集的蛋白质点进行质谱鉴定结果表明,固相配体文库技术能够提高血清低丰度蛋白质谱鉴定,如簇集蛋白、玻连蛋白、C4b结合蛋白α链、补体C4-A等。结论固相配体文库技术能有效地富集血清中低丰度蛋白,同时稀释高丰度蛋白,表明该技术有助于血清中低丰度蛋白的分离、富集与鉴定。Objective To determine the effects of solid-phase ligand libraries on enrichment of low-abundance serum protein and improve the low-abundance serum protein identification by mass spectrometer（MS）. Methods Three pooled serum samples were treated with ProteoMiner and then subjected to one-dimensional （one-D） or two-dimensional （two-D） SDS-PAGE followed by comparative image analysis. The remarkable increased protein spots on two-D map were identified by high performance liquid cbromatogrphy（HPLC） coupled on-line to ESI/MS-MS. Results The one-D and two-D gel map revealed more low-abundance protein bands and protein spots, respectively. MS identification of significantly enriched protein spots on two-D gel improved low-abundance protein identification after library beads treatment, e. g. clusterin, vitronectin, C4b-binding protein alpha chain, complement C4-A. Conclusion The technique of solid-phase ligand libraries can enrich serum low-abundance proteins whilst decrease high-abundance protein concentration, which indicates that this method is useful for enrichment of low-abundance protein and identification.

关 键 词：血清蛋白质组学 固相配体文库 簇集蛋白 高丰度蛋白 低丰度蛋白 高效液相色谱法 

分 类 号：Q46[生物学—生理学] R331.1[医药卫生—人体生理学]