Ultrasensitive detection of protein biomarker using fluorescent quantum dot probes

doi:10.3969/j.issn.1000-5641.2016.02.016

Abstract

Abstract: In this study, based on the unique properties of quantum dots (QDs) and immune magnetic beads, an fluorescent assay was built to detect low concentration carcinoembryonic antigen (CEA). The assay was carried out in a sandwich formation in which two antibodies simultaneously recognized two separate binding sites on the same target CEA. After enrichment with magnetic separator, the fluorescent intensities produced by the QDs, which were confined to the surface of the beads, could be read efficiently to quantify the concentration of the CEA. For this assay, the limit of detection was 38 pg/mL with the linear range was from 0.39 to 50 ng/mL, and it has been proved with high accuracy and repeatability in tumor marker control. With the high sensitivity, specificity and low sample consumption, this method takes a big step to the early diagnosis of diseases.

Key words: quantum dots, immunofluorescence assay, magnetic beads, tumor marker

CLC Number:

WU Si-Min, JIA Chun-Ping, LIU Li-Fen, GAO Wan-Lei, JING Feng-Xiang, Jin-Qing-Hui , ZHANG Hong-Feng, ZHAO Jian-Long-. Ultrasensitive detection of protein biomarker using fluorescent quantum dot probes[J]. Journal of East China Normal University(Natural Sc, 2016, 2016(2): 144-152.

References

［1］WAGNER M K, LI F, LI J, et al. Use of quantum dots in the development of assays for cancer biomarkers［J］. Anal Bioanal Chem, 2010, 397(8): 32133224.

［2］LI X, ASMITANANDA T, GAO L, et al. Biomarkers in the lung cancer diagnosis: A clinical perspective［J］. Neoplasma, 2012, 59(5): 500507.

［3］SUNG H J, CHO J Y. Biomarkers for the lung cancer diagnosis and their advances in proteomics［J］. BMB Rep, 2008, 41(9): 615625.

［4］SIEGEL R, MA J, ZOU Z, et al. Cancer statistics, 2014［J］. CA Cancer J Clin, 2014, 64(1): 929.

［5］MITSUHASHI N, TAKAHASHI T, SAKURAI H, et al. Establishment and characterization of a new human lung poorly differentiated adenocarcinoma cell line, GLL1, producing carcinoembryonic antigen (CEA) and CA199［J］. Lung Cancer, 1995, 12(1/2): 1324.

［6］GRUNNET M, SORENSEN J B. Carcinoembryonic antigen (CEA) as tumor marker in lung cancer［J］. Lung Cancer, 2012, 76(2): 138143.

［7］JIA C P, ZHONG X Q, HUA B, et al. NanoELISA for highly sensitive protein detection［J］. Biosens Bioelectron, 2009, 24(9): 28362841.

［8］XIANG D S, ZENG G P, HE Z K. Magnetic microparticlebased multiplexed DNA detection with biobarcoded quantum dot probes［J］. Biosens Bioelectron, 2011, 26(11): 44054410.

［9］PENG Z A, PENG X. Formation of highquality CdTe, CdSe, and CdS nanocrystals using CdO as precursor［J］. J Am Chem Soc, 2001, 123(1): 183184.

［10］PETRYAYEVA E, ALGAR W R, MEDINTZ I L. Quantum dots in bioanalysis: A review of applications across various platforms for fluorescence spectroscopy and imaging［J］. Appl Spectrosc, 2013, 67(3): 215252.

［11］KLIMOV V I. Spectral and dynamical properties of multiexcitons in semiconductor nanocrystals［J］. Annu Rev Phys Chem, 2007, 58: 635673.

［12］SMITH A M, NIE S. Semiconductor nanocrystals: Structure, properties, and band gap engineering［J］. Acc Chem Res, 2010, 43(2): 190200.

［13］YU X, XIA H S, SUN Z D, et al. Onchip dual detection of cancer biomarkers directly in serum based on selfassembled magnetic bead patterns and quantum dots［J］. Biosens Bioelectron, 2013, 41: 129136.

［14］PARK H, HWANG M P, LEE J W, et al. Harnessing immunomagnetic separation and quantum dotbased quantification capacities for the enumeration of absolute levels of biomarker［J］. Nanotechnology, 2013, 24(28): 285103.

［15］DUNBAR S A. Applications of Luminex xMAP technology for rapid, highthroughput multiplexed nucleic acid detection［J］. Clin Chim Acta, 2006, 363(12): 7182.

［16］GOKARNA A, JIN L H, HWANG J S, et al. Quantum dotbased protein micro and nanoarrays for detection of prostate cancer biomarkers［J］. Proteomics, 2008, 8(9): 18091818.

［17］CHENG X, PU X, JUN P, et al. Rapid and quantitative detection of Creactive protein using quantum dots and immunochromatographic test strips［J］. Int J Nanomedicine, 2014(9): 56195626.

［18］ZHU X, DUAN D, PUBLICOVER N G. Magnetic bead based assay for Creactive protein using quantumdot fluorescence labeling and immunoaffinity separation［J］. Analyst, 2010, 135(2): 381389.

［19］ZHOU L, OU L J, CHU X, et al. Aptamerbased rolling circle amplification: A platform for electrochemical detection of protein［J］. Anal Chem, 2007, 79(19): 74927500.

［20］WHITEAKER J R, ZHAO L, ZHANG H Y, et al. Antibodybased enrichment of peptides on magnetic beads for massspectrometrybased quantification of serum biomarkers［J］. Anal Biochem, 2007, 362(1): 4454.

［21］LIU M, JIA C, HUANG Y, et al. Highly sensitive protein detection using enzymelabeled gold nanoparticle probes［J］. Analyst, 2010, 135(2): 327331.

［22］SWIERCZEWSKA M, LIU G, LEE S, et al. Highsensitivity nanosensors for biomarker detection［J］. Chem Soc Rev, 2012, 41(7): 26412655.

［23］ESTEVETURRILLAS F A, ABADFUENTES A. Applications of quantum dots as probes in immunosensing of smallsized analytes［J］. Biosens Bioelectron, 2013, 41: 1229.

［24］WANG J, MOUNTZIARIS T J. Homogeneous immunoassays based on fluorescence emission intensity variations of zinc selenide quantum dot sensors［J］. Biosens Bioelectron, 2013, 41: 143149.