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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 investigate the effect of solid-phase ligand library on the enrichment of serum low abundance proteins and to improve the sensitivity of low abundance proteins by mass spectrometry. Methods 3 groups of mixed serum samples were treated with ProteoMiner, and one-dimensional or two-dimensional electrophoretic pattern analysis was used to compare the difference bands or protein spots in the serum profiles before and after treatment. The two-dimensional electrophoretograms Increased protein spots were identified by high performance liquid chromatography with electrospray tandem mass spectrometry. Results The one-dimensional and two-dimensional electrophoretograms showed that the solid-phase ligand library technology could display more protein bands or protein spots. Significantly enriched protein spots were identified by mass spectrometry. The results showed that solid-phase ligand library technology can improve serum low Abundance protein mass spectrometry identification, such as the cluster protein, vitronectin, C4b binding protein α chain, complement C4-A and so on. Conclusion The technique of solid-phase ligand library can effectively enrich low-abundance serum protein and high-abundance protein simultaneously, indicating that this technique is helpful for the isolation, enrichment and identification of low abundance proteins in serum.