固相配体文库降低血清高低丰度蛋白浓度差异

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...

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

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 id