Chemiluminescent optical fiber immunosensor for detection of autoantibodies to ovarian and breast cancer-associated antigens.
We report herein the development of an optical fiber based chemiluminescent immunosensor for detection of the native autoimmune response to GIPC-1, a PDZ containing protein involved in regulation of G-protein signaling. The recombinant protein GIPC-1 was expressed in bacteria, purified, refolded and conjugated to the tip of an optical fiber. A human monoclonal 27.B1 IgM isolated from a breast cancer patient, which targets the GIPC-1 protein, was used for calibration of the immunosensor and was detected down to a concentration of 30 pg/ml. We determined that the fiber-optic immunosensor had a detection limit 50 times lower than chemiluminescent ELISA, and approximately 500 times lower than colorimetric ELISA. In addition, sera from 11 ovarian cancer patients, 22 breast cancer patients and asymptomatic controls were tested for the presence of IgM anti-GIPC-1 autoantibodies in their serum using the two methods. The immunosensor assay detected 54% and 77% GIPC-1 positive sera within ovarian and breast cancer patients, respectively, as compared to chemiluminescent ELISA, which only detected 18% and 27%, respectively. We envision that this immunosensor may serve as a diagnostic tool for screening women for ovarian and breast cancer at an early stage, thus increasing their chance of survival.
Salama
Orly
O
Department of Biotechnology Engineering, The National Institute for Biotechnology in the Negev, and Soroka University Hospital, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Herrmann
Sebastien
S
Tziknovsky
Alina
A
Piura
Benjamin
B
Meirovich
Michael
M
Trakht
Ilya
I
Reed
Brent
B
Lobel
Leslie I
LI
Marks
Robert S
RS
D048868:Adaptor Proteins, Signal Transducing / Q000276:immunology; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000097:blood / Q000276:immunology*; D001323:Autoantibodies / Q000032:analysis* / Q000097:blood; D015374:Biosensing Techniques / Q000295:instrumentation*; D001943:Breast Neoplasms / Q000097:blood / Q000276:immunology*; D005260:Female; D005336:Fiber Optic Technology; D006801:Humans; D049449:Luminescence; D055100:Optical Fibers; D010051:Ovarian Neoplasms / Q000097:blood / Q000276:immunology*
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D001323:Autoantibodies; C492009:GIPC1 protein, human