Abstract
The use of colorimetric bioassays for protein detection is one of the most promising diagnostic approaches, but their relatively poor detection limits have been a critical issue. In this study, we developed an efficient colorimetric bioassay based on antibody-coated peroxidase-mimicking Au nanoparticles (Au NPs) for the detection of prostate-specific antigen (PSA), which is one of the most widely used protein biomarkers for the diagnosis of prostate and breast cancers. Anti-PSA antibody adsorption on the Au NP surface result in 65% of the peroxidasemimicking properties of Au NPs was suppressed to maximize sensitivity of the assay. Anti-PSA antibody adsorption on the Au NP was optimized at 150 µg/mL anti-PSA4 antibody for 1 h incubation, and their surface was blocked with 2% BSA. The experimental conditions of the immunoassay detection were also investigated. A highest assay value was obtained at 10 µg/mL anti-PSA1 capture antibody using 3% BSA for surface blocking and at 2.5 pM anti- PSA4-Ab coated Au NPs for 1 h incubation with PSA antigen. As a result, PSA was detected at a wide range of 0.25 to 2,500 ng/mL with a detection limit of 0.23 ng/mL. The anti-PSA-Ab coated Au NP- based assay was simple, easy to operate, and sensitive. The proposed peroxidase-mimicking anti-PSA Ab-coated Au NP-based assay could be further developed to detect other protein biomarkers.
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References
Masson, J.-F. Surface Plasmon Resonance Clinical Biosensors for Medical Diagnostics. ACS Sens. DOI https://doi.org/10.1021/acssensors.6b00763.
Madhavan, B., Yue, S., Galli, U., Rana, S., Gross, W., Müller, M., Giese, N.A., Kalthoff, H., Becker, T., Büchler, M.W. & Zöller, M. Combined evaluation of a panel of protein and miRNA serumexosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity. Int. J. Cancer DOI https://doi.org/10.1002/ijc.29324.
Xu, J., Shi, M., Chen, W., Huang, Y., Fang, L., Yao, L., Zhao, S., Chen, Z.-F. & Liang, H. A gold nanoparticle-based four-color proximity immunoassay for one-step, multiplexed detection of protein biomarkers using ribonuclease H signal amplification. Chem. Commun. (Cambridge, U. K.) DOI https://doi.org/10.1039/C7CC09404C.
Wang, Y., Dostalek, J. & Knoll, W. Magnetic nanoparticle-enhanced biosensor based on grating-coupled surface plasmon resonance. Anal. Chem. DOI https://doi.org/10.1021/ac200751s.
Nimse, S.B., Sonawane, M.D., Song, K.-S. & Kim, T. Biomarker detection technologies and future directions. Analyst DOI https://doi.org/10.1039/C5AN01790D.
Geißler, D., Stufler, S., Löhmannsröben, H.-G. & Hildebrandt, N. Six-Color Time-Resolved Förster Resonance Energy Transfer for Ultrasensitive Multiplexed Biosensing. J. Am. Chem. Soc. DOI https://doi.org/10.1021/ja310317n.
Huang, Y., Liu, X., Huang, H., Qin, J., Zhang, L., Zhao, S., Chen, Z.-F. & Liang, H. Attomolar Detection of Proteins via Cascade Strand-Displacement Amplification and Polystyrene Nanoparticle Enhancement in Fluorescence Polarization Aptasensors. Anal. Chem. 87, 8107–8114 (2015). DOI https://doi.org/10.1021/ac5041692.
Akhavan-Tafti, H., Binger, D.G., Blackwood, J.J., Chen, Y., Creager, R.S., de Silva, R., Eickholt, R.A., Gaibor, J.E., Handley, R.S., Kapsner, K.P., Lopac, S. K., Mazelis, M. E., McLernon, T. L., Mendoza, J. D., Odegaard, B. H., Reddy, S. G., Salvati, M., Schoenfelner, B. A., Shapir, N., Shelly, K. R., Todtleben, J. C., Wang, G. & Xie, W. A homogeneous chemiluminescent immunoassay method. J. Am. Chem. Soc. DOI https://doi.org/10.1021/ja312039k.
Krishnan, S., Mani, V., Wasalathanthri, D., Kumar, C.V. & Rusling, J.F. Attomolar detection of a cancer biomarker protein in serum by surface plasmon resonance using superparamagnetic particle labels. Angew. Chem. Int. Ed. DOI https://doi.org/10.1002/anie.201005607.
Munge, B.S., Coffey, A.L., Doucette, J.M., Somba, B.K., Malhotra, R., Patel, V., Gutkind, J.S. & Rusling, J.F. Nanostructured immunosensor for attomolar detection of cancer biomarker interleukin-8 using massively labeled superparamagnetic particles. Angew. Chem. Int. Ed. DOI https://doi.org/10.1002/anie.201102941.
Fang, Y., Li, Y., Zhang, M., Cui, B., Hu, Q. & Wang, L. A novel electrochemical strategy based on porous 3D graphene-starch architecture and silver deposition for ultrasensitive detection of neuron-specific enolase. Analyst DOI https://doi.org/10.1039/C8AN02230E.
Gao, Z., Hou, L., Xu, M. & Tang, D. Enhanced colorimetric immunoassay accompanying with enzyme cascade amplification strategy for ultrasensitive detection of low-abundance protein. Sci. Rep. DOI https://doi.org/10.1038/srep03966
Pham, X.-H., Hahm, E., Kim, T.H., Kim, H.-M., Lee, S.H., Lee, Y.-S., Jeong, D.H. & Jun, B.-H. Enzyme-catalyzed Ag growth on Au nanoparticle-assembled structure for highly sensitive colorimetric immunoassay. Sci. Rep. DOI https://doi.org/10.1038/s41598-018-24664-w.
de la Rica, R. & Stevens, M.M. Plasmonic ELISA for the ultrasensitive detection of disease biomarkers with the naked eye. Nat. Nanotechnol. DOI https://doi.org/10.1038/nnano.2012.186
Li, J., Fu, H.-E., Wu, L.-J., Zheng, A.-X., Chen, G.-N. & Yang, H.-H. General colorimetric detection of proteins and small molecules based on cyclic enzymatic signal amplification and hairpin aptamer probe. Anal. Chem. DOI https://doi.org/10.1021/ac3006186.
Xiao, L., Zhu, A., Xu, Q., Chen, Y., Xu, J. & Weng, J. Colorimetric biosensor for detection of cancer biomarker by Au nanoparticle-decorated Bi2Se3 nanosheets. ACS Appl. Mater. Interfaces DOI https://doi.org/10.1021/acsami.6b15750.
Song, Y., Wei, W. & Qu, X. Colorimetric biosensing using smart materials. Adv. Mater. DOI https://doi.org/10.1002/adma.201101853.
Zhang, M., Qing, G., Xiong, C., Cui, R., Pang, D.-W. & Sun, T. Dual-responsive gold nanoparticles for colorimetric recognition and testing of carbohydrates with a dispersion-dominated chromogenic process. Adv. Mater. DOI https://doi.org/10.1002/adma.201203289.
Lee, J.-S., Lytton-Jean, A.K.R., Hurst, S.J. & Mirkin, C.A. Silver nanoparticle-oligonucleotide conjugates based on DNA with triple cyclic disulfide moieties. Nano Lett. DOI https://doi.org/10.1021/nl071108g.
Gao, Z., Xu, M., Hou, L., Chen, G. & Tang, D. Magnetic bead-based reverse colorimetric immunoassay strategy for sensing biomolecules. Anal. Chem. DOI https://doi.org/10.1021/ac401433p.
Malashikhina, N., Garai-Ibabe, G. & Pavlov, V. Unconventional application of conventional enzymatic substrate: first fluorogenic immunoassay based on enzymatic formation of quantum dots. Anal. Chem. DOI https://doi.org/10.1021/ac4011342.
Babaian, R.J., Johnston, D.A., Naccarato, W., Ayala, A., Bhadkamkar, V.A. & Fritsche, H.A.H.A. Jr. The incidence of prostate cancer in a screening population with a serum prostate specific antigen between 2.5 and 4.0 ng/ml: relation to biopsy strategy. J. Urol. DOI https://doi.org/10.1016/S0022-5347(05)66519-6.
Bok, R.A. & Small, E.J. Bloodborne biomolecular markers in prostate cancer development and progression. Nat. Rev. Cancer DOI https://doi.org/10.1038/nrc951.
Mannello, F. & Gazzanelli, G. Prostate-specific antigen (PSA/hK3): a further player in the field of breast cancer diagnostics? Breast Cancer Res. DOI https://doi.org/10.1186/bcr302.
Mashkoor, F.C., Al-Asadi, J.N. & Al-Naama, L.M. Serum level of prostate-specific antigen (PSA) in women with breast cancer. Cancer Epidemiol. DOI https://doi.org/10.1016/j.canep.2013.06.009.
Nam, J.-M., Thaxton, C.S. & Mirkin, C.A. Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins. Science DOI https://doi.org/10.1126/science.1088755.
Thaxton, C.S., Elghanian, R., Thomas, A.D., Stoeva, S.I., Lee, J.-S., Smith, N.D., Schaeffer, A.J., Klocker, H., Horninger, W., Bartsch, G. & Mirkin, C.A. Nanoparticle-based bio-barcode assay redefines “undetectable” PSA and biochemical recurrence after radical prostatectomy. Proc. Natl. Acad. Sci. U. S. A. DOI https://doi.org/10.1073/pnas.0904719106.
Chang, H.J., Kang, H.M., Ko, E., Jun, B.H., Lee, H.Y., Lee, Y.S. & Jeang, D.H. PSA Detection with Femtomolar Sensitivity and a Broad Dynamic Range Using SERS Nanoprobes and an Area-Scanning Method. ACS Sens. DOI https://doi.org/10.1021/acssensors.6b00053.
Chao, C.-H., Wu, C.-S., Huang, C.-C., Liang, J.-C., Wang, H.-T., Tang, P.-T., Lin, L.-Y. & Ko, F.-H. A rapid and portable sensor based on protein-modified gold nanoparticle probes and lateral flow assay for naked eye detection of mercury ion. Microelectron. Eng. DOI https://doi.org/10.1016/j.mee.2012.03.015.
de L. M. de Morais, C., Carvalho, J.C., Sant’Anna, C., Eugênio, M., Gasparotto, L.H.S. & Lima, K.M.G. A low-cost microcontrolled photometer with one color recognition sensor for selective detection of Pb2+ using gold nanoparticles. Anal. Methods DOI https://doi.org/10.1039/C5AY01762A.
Kang, J., Zhang, Y., Li, X., Miao, L. & Wu, A. A rapid colorimetric sensor of clenbuterol based on cysteamine-modified gold nanoparticles. ACS Appl. Mater. Interfaces DOI https://doi.org/10.1021/acsami.5b09079.
Rosi, N.L. & Mirkin, C.A. Nanostructures in biodiagnostics. Chem. Rev. DOI https://doi.org/10.1021/cr030067f.
Elghanian, R., Storhoff, J.J., Mucic, R.C., Letsinger, R.L. & Mirkin, C.A. Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. Science DOI https://doi.org/10.1126/science.277.5329.1078.
Marradi, M., Chiodo, F., García, I. & Penadés, S. Glyconanoparticles as multifunctional and multimodal carbohydrate systems. Chem. Soc. Rev. DOI https://doi.org/10.1039/C2CS35420A.
Zagorovsky, K. & Chan, W.C.W. A plasmonic DNAzyme strategy for point-of-care genetic detection of infectious pathogens. Angew. Chem. Int. Ed. DOI https://doi.org/10.1002/anie.201208715.
Driskell, J.D., Jones, C.A., Tompkins, S.M. & Tripp, R.A. One-step assay for detecting influenza virus using dynamic light scattering and gold nanoparticles. Analyst DOI https://doi.org/10.1039/C1AN15303J.
Lee, C., Gaston, M.A., Weiss, A.A. & Zhang, P. Colorimetric viral detection based on sialic acid stabilized goldnanoparticles. Biosens. Bioelectron. DOI https://doi.org/10.1016/j.bios.2012.10.067.
Liu, Y., Zhang, L., Wei, W., Zhao, H., Zhou, Z., Zhang, Y. & Liu, S. Colorimetric detection of influenza A virus using antibody-functionalized gold nanoparticles. Analyst DOI https://doi.org/10.1039/C5AN00407A.
Zhang, Y., McKelvie, I.D., Cattrall, R.W. & Kolev, S.D. Colorimetric detection based on localised surface plasmon resonance of gold nanoparticles: Merits, inherent shortcomings and future prospects. Talanta DOI https://doi.org/10.1016/j.talanta.2016.02.015.
Liu, L., Hao, Y., Deng, D. & Xia, N. Nanomaterials-based colorimetric immunoassays. Nanomaterials DOI https://doi.org/10.3390/nano9030316.
Ronkainen, N.J. & Okon, S.L. Nanomaterial-based electrochemical immunosensors for clinically significant biomarkers. Materials (Basel) DOI https://doi.org/10.3390/ma7064669.
Lee, G., Eom, K., Park, J., Yang, J., Haam, S., Huh, Y.-M., Ryu, J.K., Kim, N.H., Yook, J.I., Lee, S.W., Yoon, D.S. & Kwon, T. Real-time quantitative monitoring of specific peptide cleavage by a proteinase for cancer diagnosis. Angew. Chem. Int. Ed. DOI https://doi.org/10.1002/anie.201108830.
Shi, L., De Paoli, V., Rosenzweig, N. & Rosenzweig, Z. Synthesis and application of quantum dots FRET-based protease sensors. J. Am. Chem. Soc. DOI https://doi.org/10.1021/ja063509o.
Chen, G., Xie, Y., Zhang, H., Wang, P., Cheung, H.-Y., Yang, M., Sun, H. A general colorimetric method for detecting protease activity based on peptide-induced gold nanoparticle aggregation. RSC Adv. DOI https://doi.org/10.1039/C3RA46493H.
Lee, S.H. & Jun, B.H. Silver nanoparticles: synthesis and application for nanomedicine. Int. J. Mol. Sci. DOI https://doi.org/10.3390/ijms20040865.
Park, S.Y., Lee, S.M., Kim, G.B. & Kim, Y.-P. Gold nanoparticle-based fluorescence quenching via metal coordination for assaying protease activity. Gold Bull. (Berlin, Ger.) DOI https://doi.org/10.1007/s13404-012-0070-9.
Ingram, A., Byers, L., Faulds, K., Moore, B.D. & Graham, D. SERRS-based enzymatic probes for the detection of protease activity. J. Am. Chem. Soc. DOI https://doi.org/10.1021/ja803655h.
Jun, B.H., Kim, G., Jeong, S., Noh, M.S., Pham, X.H., Kang, H., Cho, M.H., Kim, J.H., Lee, Y.S. & Jeong, D.H. Silica core-based Surface-Enhanced Raman Scattering (SERS) tag: advances in multifunctional SERS nanoprobes for bioimaging and targeting of biomarkers. Bull. Korean Chem. Soc. DOI https://doi.org/10.1002/bkcs.10179.
McVey, C., Logan, N., Thanh, N.T.K., Elliott, C. & Cao, C. Unusual switchable peroxidase-mimicking nanozyme for the determination of proteolytic biomarker. Nano Res. DOI https://doi.org/10.1007/s12274-018-2241-3.
Tseng, C.-W., Chang, H.-Y., Chang, J.-Y. & Huang, C.-C. Detection of mercury ions based on mercury-induced switching of enzyme-like activity of platinum/gold nanoparticles. Nanoscale DOI https://doi.org/10.1039/C2NR31716H.
Hvolbæk, B., Janssens, T.V.W., Clausen, B.S., Falsig, H., Christensen, C.H. & Nørskov, J.K. Catalytic activity of Au nanoparticles. Nano Today DOI https://doi.org/10.1016/S1748-0132(07)70113-5.
Deng, H.-H., Weng, S.-H., Huang, S.-L., Zhang, L.-N., Liu, A.-L., Lin, X.-H. & Chen, W. Colorimetric detection of sulfide based on target-induced shielding against the peroxidase-like activity of gold nanoparticles. Anal. Chim. Acta DOI https://doi.org/10.1016/j.aca.2014.09.023.
Zhao, D., Chen, C., Lu, L., Yang, F. & Yang, X. A label-free colorimetric sensor for sulfate based on the inhibition of peroxidase-like activity of cysteamine-modified gold nanoparticles. Sens. Actuators, B DOI https://doi.org/10.1016/j.snb.2015.04.010.
Hizir, M.S., Top, M., Balcioglu, M., Rana, M., Robertson, N.M., Shen, F., Sheng, J. & Yigit, M.V. Multiplexed activity of perAuxidase: DNA-capped AuNPs act as adjustable Peroxidase. Anal. Chem. DOI https://doi.org/10.1021/acs.analchem.5b03926.
Shah, J., Purohit, R., Singh, R., Karakoti, A.S. & Singh, S. ATP-enhanced peroxidase-like activity of gold nanoparticles. J. Colloid Interface Sci. DOI https://doi.org/10.1016/j.jcis.2015.06.015.
Turkevich, J., Stevenson, P.C. & Hillier, J. A study of the nucleation and growth processes in the synthesis of colloidal gold. Discuss. Faraday Soc. DOI https://doi.org/10.1039/DF9511100055.
Haiss, W., Thanh, N.T.K., Aveyard, J. & Fernig, D.G. Determination of size and concentration of gold nanoparticles from UV-vis spectra. Anal. Chem. DOI https://doi.org/10.1021/ac0702084.
Vignati, G., Chiecchio, A., Osnaghi, B., Giovanelli, L. & Meloncelli, C. Different biological matrices (serum and plasma) utilization in consolidation processes: evaluation of seven Access immunoassays. Clin. Chem. Lab. Med. DOI https://doi.org/10.1515/CCLM.2008.032
Liu, Y., Zhang, Z., Yu, J., Xie, J. & Li, C.M. A concentration-dependent multicolor conversion strategy for ultrasensitive colorimetric immunoassay with the naked eye. Anal. Chim. Acta DOI https://doi.org/10.1016/j.aca.2017.01.034.
Suaifan, G.A.R.Y., Esseghaier, C., Ng, A. & Zourob, M. Ultra-rapid colorimetric assay for protease detection using magnetic nanoparticle-based biosensors. Analyst DOI https://doi.org/10.1039/C3AN36881E.
Fu, G., Sanjay, S.T. & Li, X. Cost-effective and sensitive colorimetric immunosensing using an iron oxide-to-Prussian blue nanoparticle conversion strategy. Analyst DOI https://doi.org/10.1039/C6AN00254D.
Hahn, J., Kim, E., You, Y. & Choi, Y.J. Colorimetric switchable linker-based bioassay for ultrasensitive detection of prostate-specific antigen as a cancer biomarker. Analyst DOI https://doi.org/10.1039/C9AN00552H.
Lai, W., Tang, D., Zhuang, J., Chen, G. & Yang, H. Magnetic bead-based enzyme-chromogenic substrate system for ultrasensitive colorimetric immunoassay accompanying cascade reaction for enzymatic formation of squaric acid-iron(III) chelate. Anal. Chem. DOI https://doi.org/10.1021/ac500738a.
Cao, C., Li, X., Lee, J. & Sim, S.J. Homogenous growth of gold nanocrystals for quantification of PSA protein biomarker. Biosens. Bioelectron. DOI https://doi.org/10.1016/j.bios.2008.07.046.
Acknowledgements
This work was supported by the KU Research Professor Program of Konkuk University and funded by Ministry of Science, ICT and Future Planning (NRF-2016M3A9B6918892), and the Korean Health Technology R&D Project, Ministry of Health & Welfare (HI17C1264).
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Pham, XH., Hahm, E., Huynh, KH. et al. Sensitive Colorimetric Detection of Prostate Specific Antigen Using a Peroxidase-Mimicking Anti-PSA Antibody Coated Au Nanoparticle. BioChip J 14, 158–168 (2020). https://doi.org/10.1007/s13206-019-4204-5
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DOI: https://doi.org/10.1007/s13206-019-4204-5