Applying Proteomics Technology In Screening And Identifying Tumor Antigens Of Ovarian Carcinoma

BACKGROUD&OBJECTIVE: Ovarian carcinoma is the major cause which leads to death in women in malignancies. Early diagnosis impacts the prognosis of the ovarian carcinoma patients. Screening tumor antigens plays a key role in diagnosis of ovarian carcinoma in the early stage, offers new therapeutic targets and designs vaccines in immunotherapy for ovarian carcinoma. With the concept of proteome appeared in 1995, proteomics became rapidly a new front area of life science, proteomic technology also developed swift and palyed an important role in various research areas. Several technologies such as phage antibody library, ribosome display and serological analysis of recombinant cDNA expression libraries (SEREX) have been applied widely. In the present study, proteomic technologies were applied to screen and identify tumor antigens of ovarian carcinoma, then to establish a new methodological entity.METHODS: The initial treatment of ovarian carcinoma patients' sera were collected; Western blot was used to find out the obviously different serum in ovarian carcinoma patients; SKOV3 lysate was added with the patient's serum; immunoprecipitation was used to enrich antigens; antigens were eluted, then SDS-PAGE. The different protein bands were gained compared with normal serum immunoprecipitates, identified by mass spectrometry andanalysed with bioinformatics.RESULTS: Two sera were the obviously different ones with Western blot; with immunoprecipitation and SDS-PAGE, three different protein bands of the patients appeared in the position of about 66kDa and 42kDa compared with normal serum immunoprecipitates, and identified by mass spectrometry. Then through bioinformatics analysis, heat shock protein 70(HSP70), cytokeratin 9, cytokeratin 7 and KIAA1157 were identified in the three different protein bands respectively.CONCLUSIONS: Special ovarian carcinoma patient sera were gained by primary screening with Western blot, then combined immunoprecipitation and mass spectrometry of proteomic technologies to enrich antigens and identify the different protein bands, which is an effectively methodological entity for screening and identifying tumor antigens.