Protein microarrays and quantum dot probes for early cancer detection.
We describe here a novel approach for detection of cancer markers using quantum dot protein microarrays. Both relatively new technologies; quantum dots and protein microarrays, offer very unique features that together allow detection of cancer markers in biological specimens (serum, plasma, body fluids) at pg/ml concentration. Quantum dots offer remarkable photostability and brightness. They do not exhibit photobleaching common to organic fluorophores. Moreover, the high emission amplitude for QDs results in a marked improvement in the signal to noise ratio of the final image. Protein microarrays allow highly parallel quantitation of specific proteins in a rapid, low-cost and low sample volume format. Furthermore the multiplexed assay enables detection of many proteins at once in one sample, making it a powerful tool for biomarker analysis and early cancer diagnostics. In a series of multiplexing experiments we investigated ability of the platform to detect six different cytokines in protein solution. We were able to detect TNF-alpha, IL-8, IL-6, MIP-1beta, IL-13 and IL-1beta down to picomolar concentration, demonstrating high sensitivity of the investigated detection system. We have also constructed and investigated two different models of quantum dot probes. One by conjugation of nanocrystals to antibody specific to the selected marker--IL-10, and the second by use of streptavidin coated quantum dots and biotinylated detector antibody. Comparison of those two models showed better performance of streptavidin QD-biotinylated detector antibody model. Data quantitated using custom designed computer program (CDAS) show that proposed methodology allows monitoring of changes in biomarker concentration in physiological range.
Zajac
Aleksandra
A
Division of Cancer Prevention and Control, H. Lee Moffitt Cancer Center and Research Institute, University of South Florida, 12902 Magnolia Drive, Tampa, FL 33612, USA.
Song
Dansheng
D
Qian
Wei
W
Zhukov
Tatyana
T
D000818:Animals; D000911:Antibodies, Monoclonal; D016207:Cytokines / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D015336:Molecular Probe Techniques; D015335:Molecular Probes; D009369:Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D040081:Protein Array Analysis; D045663:Quantum Dots; D051381:Rats
D000911:Antibodies, Monoclonal; D016207:Cytokines; D015335:Molecular Probes