Comparison of 1-D and 2-D LC MS/MS methods for proteomic analysis of human serum
Comparison of 1-D and 2-D LC MS/MS methods for proteomic analysis of human serum
复制标题
DOI：
10.1002/elps.200800630
复制
发表时间：
2009-04-01
期刊：
影响因子：
2.9
通讯作者：
Gebler, John C.
中科院分区：
文献类型：
--
作者：
Gilar, Martin;Olivova, Petra;Gebler, John C.
1-D和2-D LC方法用于未耗尽的人血清的蛋白质组分析。用强阳离子交换（SCX）-RP色谱法或用RP-RP 2-D LC方法在2-D LC中进行肽的分离。使用数据独立的采集方法通过MS/MS鉴定肽。肽保留预测模型用于突出潜在的假阳性肽鉴定。当应用选定的数据过滤，我们确定了52个蛋白质的基础上316肽血清中的1-D LC设置。在RP-RP和SCX-RP 2-D LC中分别鉴定了184个蛋白质/1036个肽和142个蛋白质/905个肽。这两个2-D LC方法的蛋白质组学分析的性能进行了严格的比较。
1-D and 2-D LC methods were utilized for proteome analysis of undepleted human serum. Separation of peptides in 2-D LC was performed either with strong cation exchange (SCX)-RP chromatography or with an RP-RP 2-D LC approach. Peptides were identified by MS/MS using a data-independent acquisition approach. A peptide retention prediction model was used to highlight the potential false-positive peptide identifications. When applying selected data filtration, we identified 52 proteins based on 316 peptides in serum in 1-D LC setup. One hundred and eighty-four proteins/1036 peptides and 142 proteins/905 peptides were identified in RP-RP and SCX-RP 2-D LC, respectively. The performance of both 2-D LC methods for proteomic analysis is critically compared.