Quantum dots-based reverse phase protein microarray.
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 (R(2)=0.986). This suggests that Qdots can be used for protein quantification in microarray format, using the platform presented here.
Shingyoji
Masato
M
Life Sciences Division, Lawrence Berkley National Laboratory, University of California at Berkeley, MS 977R0225A, 1 Cyclotron Road, Berkeley, CA 94720, USA.
Gerion
Daniele
D
Pinkel
Dan
D
Gray
Joe W
JW
Chen
Fanqing
F