Detection of eight antibodies in cancer patients' sera against proteins derived from the adenocarcinoma A549 cell line using proteomics-based analysis.
To screen cancer for specific autoantibodies, we applied the approach established by Brichory et al., who reported annexins I and II as specific antigens. Solubilized proteins from a cancer cell line (A549) were separated using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), followed by Western blotting (WB) analysis, in which the sera of individual patients were tested for primary antibodies. We found 11 positive spots on PVDF membrane using a WB/enhanced chemiluminescence detection Kit, and identified eight proteins, such as alpha-enolase, inosine-5'-monophosphate dehydrogenase, aldehyde dehydrogenase, 3-phosphoglycerate dehydrogenase, 3-oxoacid CoA transferase, chaperonin, peroxiredoxin 6 and triosephosphate isomerase, that reacted with these antibodies in patients' sera using MALDI-TOF/TOF. All eight antibodies were not detected in the sera derived from lung tuberculosis and healthy controls.
Nakanishi
Toyofumi
T
Department of Clinical Pathology, Osaka Medical College, Takatsuki-city 569-8686, Japan.
Takeuchi
Toru
T
Ueda
Kazuhito
K
Murao
Hitoshi
H
Shimizu
Akira
A
D000230:Adenocarcinoma / Q000276:immunology / Q000473:pathology; D000912:Antibodies, Neoplasm / Q000032:analysis* / Q000097:blood; D001323:Autoantibodies / Q000032:analysis / Q000097:blood; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology / Q000473:pathology; D009363:Neoplasm Proteins / Q000276:immunology*; D010751:Phosphopyruvate Hydratase / Q000276:immunology; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000912:Antibodies, Neoplasm; D001323:Autoantibodies; D009363:Neoplasm Proteins; D010751:Phosphopyruvate Hydratase