Quantum dots-based reverse phase protein microarray

Masato Shingyoji, Daniele Gerion, Dan Pinkel, Joe Gray, Fanqing Chen

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)472-478
Number of pages7
JournalTalanta
Volume67
Issue number3
DOIs
StatePublished - Sep 15 2005
Externally publishedYes

Fingerprint

Quantum Dots
Protein Array Analysis
Microarrays
Semiconductor quantum dots
Horseradish Peroxidase
Proteins
Dilution
Fluorescent Dyes
Fluorophores
Nanoparticles
Quantum yield
Nanocrystals
Fluorescence
Assays
Technology
Wavelength

Keywords

  • Bioconjugation
  • DNA-PK
  • Protein microarray
  • Quantum dots
  • Reverse phase protein lysate microarray
  • Tyramide signal amplification

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy

Cite this

Shingyoji, M., Gerion, D., Pinkel, D., Gray, J., & Chen, F. (2005). Quantum dots-based reverse phase protein microarray. Talanta, 67(3), 472-478. https://doi.org/10.1016/j.talanta.2005.06.064

Quantum dots-based reverse phase protein microarray. / Shingyoji, Masato; Gerion, Daniele; Pinkel, Dan; Gray, Joe; Chen, Fanqing.

In: Talanta, Vol. 67, No. 3, 15.09.2005, p. 472-478.

Research output: Contribution to journalArticle

Shingyoji, M, Gerion, D, Pinkel, D, Gray, J & Chen, F 2005, 'Quantum dots-based reverse phase protein microarray', Talanta, vol. 67, no. 3, pp. 472-478. https://doi.org/10.1016/j.talanta.2005.06.064
Shingyoji, Masato ; Gerion, Daniele ; Pinkel, Dan ; Gray, Joe ; Chen, Fanqing. / Quantum dots-based reverse phase protein microarray. In: Talanta. 2005 ; Vol. 67, No. 3. pp. 472-478.
@article{baa51acf0bf2424c98fd94e9dca22325,
title = "Quantum dots-based reverse phase protein microarray",
abstract = "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.",
keywords = "Bioconjugation, DNA-PK, Protein microarray, Quantum dots, Reverse phase protein lysate microarray, Tyramide signal amplification",
author = "Masato Shingyoji and Daniele Gerion and Dan Pinkel and Joe Gray and Fanqing Chen",
year = "2005",
month = "9",
day = "15",
doi = "10.1016/j.talanta.2005.06.064",
language = "English (US)",
volume = "67",
pages = "472--478",
journal = "Talanta",
issn = "0039-9140",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Quantum dots-based reverse phase protein microarray

AU - Shingyoji, Masato

AU - Gerion, Daniele

AU - Pinkel, Dan

AU - Gray, Joe

AU - Chen, Fanqing

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

UR - http://www.scopus.com/inward/record.url?scp=24144472343&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=24144472343&partnerID=8YFLogxK

U2 - 10.1016/j.talanta.2005.06.064

DO - 10.1016/j.talanta.2005.06.064

M3 - Article

C2 - 18970191

AN - SCOPUS:24144472343

VL - 67

SP - 472

EP - 478

JO - Talanta

JF - Talanta

SN - 0039-9140

IS - 3

ER -