Bouras G.,7 Daigaku cho |
Nakanishi T.,Osaka Medical College |
Fujita Y.,7 Daigaku cho |
Tsunemi S.,7 Daigaku cho |
And 2 more authors.
Analytical and Bioanalytical Chemistry | Year: 2012
An experiment was conducted for the identification of β-tubulin as a common immunogen in gastrointestinal malignancy by mass spectrometry of colorectal cancer proteome. To determine the percentage of cancer patients and healthy subjects with antibodies against β-tubulin, commercially available recombinant β-tubulin was placed directly onto pieces of nitrocellulose membrane at 60 ng/2 μL. The membrane pieces were blocked, washed, and reacted with patient sera and horseradish peroxidase-conjugated mouse anti-human immunoglobulin IgG at 1:200 and 1:2,500 dilutions, respectively, followed by immunodetection by chemiluminescence. Disease stage, tumor differentiation, and vascular and lymphatic invasion were compared between the antibody-positive and antibody-negative groups. Out of several spots found in cancer patients, one spot located just above 50 kDa with a pI of around 5 was found to occur most frequently and selected for mass spectrometry.