Analysis of proteomic components of sera from patients with hepatocellular carcinomas by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time of flying mass spectrometry / 中华病理学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the proteomic components of the sera from the patients with hepatocellular carcinomas (HCC), in search of the diagnostic markers of HCC.</p><p><b>METHODS</b>Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2DE), silver staining, PDQuest 2DE analysis software, peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) and SWISS-PROT database searching, were used to separate and identify the proteome of the sera from the patients with HCC.</p><p><b>RESULTS</b>The 2DE images were analysed by PDQuest 2DE software. The average matching rate was 70.2%. In IEF direction, the average deviation was (1.02 +/- 0.22) mm, in SDS-PAGE direction, the deviation was (0.97 +/- 0.14) mm. The twenty-three different protein spots were incised from silver staining gel and digested in-gel by TPCK trypsin. 15 peptide mass fingerprints (PMF) maps were obtained by MOLDI-TOF-MS. The typical peptide masses were searched in the SWISS-PROT database using PeptIdent software.</p><p><b>CONCLUSIONS</b>Good reproducibility could be obtained by applying immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2DE) to separate and identify the proteome in serum. There is still the problem of efficiently removing a higher level of proteins and lipids from the serum. Identification by MOLDI-TOF-MS peptide mass fingerprint provides useful information for screening diagnostic markers of human HCC.</p>