Gilar Martin, Olivova Petra, Chakraborty Asish B, Jaworski Aleksander, Geromanos Scott J, Gebler John C
Waters Corporation, Milford, MA 01757, USA.
Electrophoresis. 2009 Apr;30(7):1157-67. doi: 10.1002/elps.200800630.
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.
一维液相色谱（1-D LC）和二维液相色谱（2-D LC）方法用于未耗尽人血清的蛋白质组分析。二维液相色谱中肽段的分离采用强阳离子交换（SCX）-反相（RP）色谱法或反相-反相二维液相色谱法。使用数据非依赖采集方法通过串联质谱（MS/MS）鉴定肽段。使用肽保留预测模型突出潜在的假阳性肽段鉴定结果。应用选定的数据过滤方法时，在一维液相色谱设置中，基于血清中的316个肽段鉴定出52种蛋白质。在反相-反相二维液相色谱和强阳离子交换-反相二维液相色谱中分别鉴定出184种蛋白质/1036个肽段和142种蛋白质/905个肽段。对两种二维液相色谱方法用于蛋白质组分析的性能进行了严格比较。
