Application of Carbon Nanotubes Coated on Silicon Wafer Combined with Immuno-PCR on the Early Diagnosis of Breast Cancer Recurrence
Date Issued
2010
Date
2010
Author(s)
Chang, Chun-Wei
Abstract
Breast cancer is the most commonly diagnosed cancer for women population in Taiwan. If breast cancer recurrence happens, there would be very high possibility for distant metastasis, which could not be cured and lead to a worse prognosis. Breast cancer is commonly diagnosed by breast images, which is not easy to detect the tumor due to technical limitations. Also, it accompanies with radioactive rays and may hazard patients’ health. In this study, we utilized immuno-PCR as a tool for diagnostics. Our purpose was to develop a precise (due to the specificity between antigen and antibody) and sensitive (due to the amplification of DNA by polymerase chain reaction) method to diagnose breast cancer recurrence from patients’ serum. Furthermore, we hoped that patients could be treated properly as early as possible for longer survival periods.
First, silicon wafer, being treated by hydrofluoric acid, was used as the substrate, with bundled carbon nanotubes (CNT) coated on it. Then, we used HNO3 / H2SO4 solution to do the surface modification and carboxyl groups would be grafted on the tips of CNT. Next, polyoxyethylene bis-amine (PEG bis-amine) would be grafted and capture antibodies (anti human CA 15-3 antibody) were conjugated. Water contact angle measurements, scanning electron microscope (SEM), thermal analysis (TA), Fourier transform infrared spectrometer (FTIR), Raman scattering spectrometer and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) would be used to prove that the surface modification was successful. The capture antibodies conjugated on the CNT were used to detect antigens (CA 15-3). Biotinylated detection antibodies (secondary antibodies) were further bound to the antigens. Free streptavidin was used to link biotinylated DNA to the biotinylated detection antibodies. The biotinylated DNA was amplified by PCR, and analyzed by agarose gel electrophoresis and NanoDrop.
According to the experimental results, the substrate, PEG bis-amine grafted CNT coated on the hydrophobic surface of silicon wafer, could prevent non-specific binding of protein and DNA and then reduce background noise in immunoassays. By using the parameters in this study, the sensitivity for carbohydrate antigen 15-3 (CA 15-3) detection by using our immuno-PCR system was 500-5000 folds higher than ELISA.
Subjects
breast cancer
carbon nanotubes
CA 15-3
immuno-PCR
ELISA
SDGs
Type
thesis
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