TY - JOUR
T1 - Quantum dots-based reverse phase protein microarray
AU - Shingyoji, Masato
AU - Gerion, Daniele
AU - Pinkel, Dan
AU - Gray, Joe W.
AU - Chen, Fanqing
N1 - Funding Information:
We thank Mr. Kevin Peet for excellent administrative support, we also thank Johnathan Zhang, Shota Yamamoto, and Dustin Dean for excellent technical support. We thank Drs. Steve Yannone and Benjamin P. Chen for providing antibody to target proteins, Drs. N. Zaitseva and G. Galli for their support, and Prof. A.P. Alivisatos for access to his laboratory. This work was supported by NIH Grant R21CA95393-01, by NASA grant NNA04CA75I, by Department of Energy grant to F. Chen and by NIH P50 grant CA112970 to J.W. Gray. This work was performed under the auspices of the U.S. Department of Energy, at the University of California/Lawrence Livermore National Laboratory under contract no. W-7405-Eng-48, and at the University of California/Lawrence Berkeley National Laboratory under contract no. DE-AC03-76SF00098.
PY - 2005/9/15
Y1 - 2005/9/15
N2 - CdSe nanocrystals, also called quantum dots (Qdots) are a novel class of fluorophores, which have a diameter of a few nanometers and possess high quantum yield, tunable emission wavelength and photostability. They are an attractive alternative to conventional fluorescent dyes. Quantum dots can be silanized to be soluble in aqueous solution under biological conditions, and thus be used in bio-detection. In this study, we established a novel Qdot-based technology platform that can perform accurate and reproducible quantification of protein concentration in a crude cell lysate background. Protein lysates have been spiked with a target protein, and a dilution series of the cell lysate with a dynamic range of three orders of magnitude has been used for this proof-of-concept study. The dilution series has been spotted in microarray format, and protein detection has been achieved with a sensitivity that is at least comparable to standard commercial assays, which are based on horseradish peroxidase (HRP)-catalyzed diaminobenzidine (DAB) chromogenesis. The data obtained through the Qdot method has shown a close linear correlation between relative fluorescence unit and relative protein concentration. The Qdot results are in almost complete agreement with data we obtained with the well-established HRP-DAB colorimetric array (R2 = 0.986). This suggests that Qdots can be used for protein quantification in microarray format, using the platform presented here. Published by Elsevier B.V.
AB - CdSe nanocrystals, also called quantum dots (Qdots) are a novel class of fluorophores, which have a diameter of a few nanometers and possess high quantum yield, tunable emission wavelength and photostability. They are an attractive alternative to conventional fluorescent dyes. Quantum dots can be silanized to be soluble in aqueous solution under biological conditions, and thus be used in bio-detection. In this study, we established a novel Qdot-based technology platform that can perform accurate and reproducible quantification of protein concentration in a crude cell lysate background. Protein lysates have been spiked with a target protein, and a dilution series of the cell lysate with a dynamic range of three orders of magnitude has been used for this proof-of-concept study. The dilution series has been spotted in microarray format, and protein detection has been achieved with a sensitivity that is at least comparable to standard commercial assays, which are based on horseradish peroxidase (HRP)-catalyzed diaminobenzidine (DAB) chromogenesis. The data obtained through the Qdot method has shown a close linear correlation between relative fluorescence unit and relative protein concentration. The Qdot results are in almost complete agreement with data we obtained with the well-established HRP-DAB colorimetric array (R2 = 0.986). This suggests that Qdots can be used for protein quantification in microarray format, using the platform presented here. Published by Elsevier B.V.
KW - Bioconjugation
KW - DNA-PK
KW - Protein microarray
KW - Quantum dots
KW - Reverse phase protein lysate microarray
KW - Tyramide signal amplification
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U2 - 10.1016/j.talanta.2005.06.064
DO - 10.1016/j.talanta.2005.06.064
M3 - Article
C2 - 18970191
AN - SCOPUS:24144472343
SN - 0039-9140
VL - 67
SP - 472
EP - 478
JO - Talanta
JF - Talanta
IS - 3
ER -