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Detection of autoantibodies against cyclophilin A and triosephosphate isomerase in sera from breast cancer patients by proteomic analysis
Author(s) -
Tamesa Michiko Sato,
Kuramitsu Yasuhiro,
Fujimoto Masanori,
Maeda Noriko,
Nagashima Yukiko,
Tanaka Toshiyuki,
Yamamoto Shigeru,
Oka Masaaki,
Nakamura Kazuyuki
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200800675
Subject(s) - autoantibody , triosephosphate isomerase , cyclophilin a , breast cancer , cancer , antibody , microbiology and biotechnology , isomerase , blot , biology , cyclophilin , enzyme , chemistry , pathology , biochemistry , medicine , immunology , gene , genetics
Much interest is presently being shown toward identifying markers for the detection of breast cancer. To detect autoantibodies that could represent diagnostic markers for breast cancer, we comprehensively analyzed serum autoantibodies showing immunoreactivity to proteins in tumor tissues of breast cancer. Tumor tissues were obtained from 40 patients with breast cancer, along with sera from 30 other patients with breast cancer and 22 healthy donors. Proteins from tumor tissues were separated by 2‐DE. After blotting onto PVDF membranes, tissue proteins were immunoblotted with sera from patients or healthy donors. By comparing each immunoblot pattern, three immunoreactive spots displayed stronger staining intensity with patient sera than with sera from healthy donors. The matched protein spots on 2‐DE gels were digested and used for LC‐MS/MS analysis, and identified as cyclophilin A (peptidyl‐prolyl cis ‐ trans isomerase A), triosephosphate isomerase and ubiquitin‐conjugating enzyme E2N. Immunoblot analysis was then performed using commercially available purified proteins, confirming the specificity of anti‐cyclophilin A and anti‐triosephosphate isomerase antibodies in sera from patients.