中文摘要：

利用肽配体库技术（CPLL）研究了蛋清中的低丰度蛋白质组。考察了盐浓度和洗脱顺序差异对CPLL富集效率的影响。结果表明：盐浓度越高,蛋清蛋白质溶液的离子相互作用越强。利用洗脱溶液解离差异,采用先HOS（有机水溶液）后尿素CHAPS（3-[3-（胆酰胺丙基）二甲氨基]-1-丙磺酸内盐）的洗脱顺序,溶菌酶能够得到高效分离。保持蛋清的原p H 8.8,25 mmol/L KH2PO4,150 mmol/L Na Cl,尿素CHAPS洗脱,是CPLL提供高效富集的前提。通过分析CPLL富集前后的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDSPAGE）条带差异,采用线性离子阱（LTQ）质谱在蛋清中检出45种低丰度蛋白质,其中两种未见报道。对蛋清中低丰度蛋白质亚细胞的定位分析发现,蛋清蛋白质的分泌蛋白居多。研究结果表明,CPLL技术能够实现蛋清中低丰度蛋白质的高效富集,分析深度达到亚细胞水平。

英文摘要：

Low-abundance proteome of chicken egg white were studied by using combinatorial peptide ligand libraries（ CPLL） and the linear ion trap quadrupole（ LTQ） mass spectrometry. The effect of salt concentration and elution order on the enrichment efficiency of CPLL was investigated. The results showed that the higher salt concentration,the stronger interaction between the egg white protein solutions,which effected CPLL enrichment efficiency significantly. The lysozyme could be separated effectively in the order of urea CHAPS（ 3-[3-cholamidopropyldimethylammonio]-1-propansulfonate）elution first,HOS（ hydro-organic solution） the second elution. High effective enrichmental conditions of CPLL were chicken egg white with 150 mmol / L Na Cl,25 mmol / L KH2PO4,p H 8. 8,urea CHAPS elution. Through the analysis of the SDS- PAGE band difference with 60 μg protein loaded after CPLL enrichment,45 species of low abundance proteins from chicken egg white were identified by LTQ mass spectrometry,of which two species were not reported yet. The result of subcellular localization analysis showed that egg white protein secreted protein was 19 species,which was in the majority. The number of unknown subcellular location of proteins from chicken egg white was taken the second place. This paper shows that the CPLL technology is rapid,simple and sensitive,and is suitable for the determination and confirmation of the low-abundance protein in chicken egg white,that the depth of which reach the sub cellular level.