Abstract
A simple and sensitive electrogenerated chemiluminescence biosensor was developed to monitor matrix metalloproteinase 2 (MMP-2) by employing a specific peptide (CGPLGVRGK) as a molecular recognition substrate. Bis(2,2′-bipyridine)-4′-methyl-4-carboxybipyridine-ruthenium N-succinimidyl ester-bis(hexafluorophosphate) (Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 (Ru1) was used as ECL-emitting species and covalently labeled onto the peptide through NH2-containing lysine on the peptide via acylation reaction to form Ru1-peptide as an ECL probe. An ECL peptide-based biosensor was fabricated by self-assembling the ECL probe onto the surface of gold electrode. MMP-2 can specifically cleave the Ru1-peptide on the electrode surface, which led the partly Ru1-peptide to leave the electrode surface and resulted in the decrease of the ECL intensity obtained from the resulted electrode in 0.1 M phosphate-buffered saline (pH 7.4) containing tri-n-propylamine. The decreased ECL intensity was piecewise linear to the concentration of MMP-2 in the range from 1 to 500 ng/mL. Moreover, the ECL biosensor is successfully applied to detection of MMP-2 secreted by living cell, such as HeLa cells. Additionally, the biosensor was also applied to the evaluation of matrix metalloproteinase inhibitors. The strategy presented here is promising for other disease-related matrix metalloproteinase assay and matrix metalloproteinase inhibitor profiling with sensitivity and simplicity.
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Alouini MA, Moustoifa EF, Rubio SA, Bartegi A, Berthelot T, Déléris G. Design, characterization, and evaluation of peptide arrays allowing the direct monitoring of MMP activities. Anal Bioanal Chem. 2012;403:185–94.
Yang J, Zhang Z, Lin J, Lu J, Liu B, Zeng S, et al. Detection of MMP activity in living cells by a genetically encoded surface-displayed FRET sensor. Biochim Biophys Acta. 2007;1773:400–7.
Funovic M, Weissleder R, Tung C-H. Protease sensors for bioimaging. Anal Bioanal Chem. 2003;377:956–63.
Sheen-Chen SM, Chen HS, Eng HL, Sheen CC, Chen WJ. Serum levels of matrix metalloproteinase 2 in patients with breast cancer. Cancer Lett. 2001;173:79–82.
Vasala K, Paakko P, Turpeenniemi-Hujanen T. Matrix metalloproteinase-2 immunoreactive protein as a prognostic marker in bladder cancer. Urology. 2003;62:952–7.
Shi MM, Yu B, Gao HG, Mu JW, Ji CW. Matrix metalloproteinase 2 overexpression and prognosis in colorectal cancer: a meta-analysis. Mol Biol Rep. 2013;40:617–23.
Gupta B, Mai K, Lowe SB, Wakefield D, Nick Girolamo D, Gaus K, et al. Ultrasensitive and specific measurement of protease activity using functionalized photonic crystals. Anal Chem. 2015;87:9946–53.
Parsons SL, Watson SA, Brown PD, Collins HM, Steele RJC. Matrix metalloproteinases. Br J Surg. 1997;84:160–6.
Parsons SL, Watson SA, Collins HM, Griffin NR, Clarke PA, Steele RJC. Gelatinase (MMP-2 and -9) expression in gastrointestinal malignancy. Br J Cancer. 1998;78:1495–502.
Philip S, Bulbule A, Kundu GC. Osteopontin stimulates tumor growth and activation of promatrix metalloproteinase-2 through nuclear factor-b-mediated induction of membrane type 1 matrix metalloproteinase in murine melanoma cells. J Biol Chem. 2001;276:44926–35.
Patel S, Sumitra G, Koner BC, Saxena A. Role of serum matrix metalloproteinase-2 and -9 to predict breast cancer progression. Clin Biochem. 2011;44:869–72.
Qi H, Li M, Dong M, Ruan SP, Gao Q, Zhang C. Electrogenerated chemiluminescence peptide-based biosensor for the determination of prostate-specific antigen based on target-induced cleavage of peptide. Anal Chem. 2014;86:1372–9.
Gao L, Mbonu N, Cao L, Gao D. Label-free colorimetric detection of gelatinases on nanoporous silicon photonic films. Anal Chem. 2008;80:1468–73.
Moss ML, Koller G, Bartsch JW, Rakow S, Schlomann U, Rasmussen FH. A colorimetric-based amplification system for proteinases including MMP2 and ADAM8. Anal Biochem. 2015;484:75–81.
Wang Z, Li X, Feng D, Li L, Shi W, Ma H. Poly(m-phenylenediamine)-based fluorescent nanoprobe for ultrasensitive detection of matrix metalloproteinase 2. Anal Chem. 2014;86:7719–25.
Wang Y, Shen P, Li C, Wang Y, Liu Z. Upconversion fluorescence resonance energy transfer based biosensor for ultrasensitive detection of matrix metalloproteinase-2 in blood. Anal Chem. 2012;84:1466–73.
Mok H, Bae KH, Ahn CH, Park TG. PEGylated and MMP-2 specifically dePEGylated quantum dots: comparative evaluation of cellular uptake. Langmuir. 2009;25:1645–50.
Lee S, Cha EJ, Park K, Lee SY, Hong JK, Sun IC, et al. A near-infrared-fluorescence-quenched gold-nanoparticle imaging probe for in vivo drug screening and protease activity determination. Angew Chem. 2008;120:2846–9.
Yang G, Li L, Rana RK, Zhu J. Assembled gold nanoparticles on nitrogen-doped graphene for ultrasensitive electrochemical detection of matrix metalloproteinase 2. Carbon. 2013;61:357–66.
Zheng T, Zhang R, Zhang QF, Tan T, Zhang K, Zhu J, et al. Ultrasensitive dual-channel detection of matrix metalloproteinase-2 in human serum using gold-quantum dot core–satellite nanoprobes. Chem Commun. 2013;49:7881–3.
Jing P, Yi H, Xue S, Yuan R, Xu W. A ‘signal on-off’ electrochemical peptide biosensor for matrix metalloproteinase 2 based on target induced cleavage of a peptide. RSC Adv. 2015;5:65725–30.
Bard AJ. Electrogenerated chemiluminescence. New York: Marcel Dekker; 2004.
Richter MM. Electrochemiluminescence (ECL). Chem Rev. 2004;104:3003–36.
Miao W. Electrogenerated chemiluminescence and its biorelated applications. Chem Rev. 2008;108:2506–53.
Marquette CA, Blum LJ. Electro-chemiluminescent biosensing. Anal Bioanal Chem. 2008;390:155–68.
Forster RJ, Bertoncello P, Keyes TE. Electrogenerated chemiluminescence. Annu Rev Anal Chem. 2009;2:359–85.
Hu L, Xu G. Applications and trends in electrochemiluminescence. Chem Soc Rev. 2010;39:3275–304.
Rampazzo E, Bonacchi S, Genovese D, Juris R, Marcaccio M, Montalti M, et al. Nanoparticles in metal complexes-based electrogenerated chemiluminescence for highly sensitive applications. Coord Chem Rev. 2012;256:1664–81.
Bertoncello P, Stewart AJ, Dennany L. Analytical applications of nanomaterials in electrogenerated chemiluminescence. Anal Bioanal Chem. 2014;406:5573–87.
Deng S, Lei J, Huang Y, Cheng Y, Ju H. Electrochemiluminescent quenching of quantum dots for ultrasensitive immunoassay through oxygen reduction catalyzed by nitrogen-doped graphene-supported hemin. Anal Chem. 2013;85:5390–6.
Wang H, Yuan Y, Chai Y, Yuan R. Sandwiched electrochemiluminescent peptide biosensor for the detection of prognostic indicator in early-stage cancer based on hollow, magnetic, and self-enhanced nanosheets. Small. 2015;11:3703–9.
Castor KJ, Metera KL, Tefashe UM, Serpell CJ, Mauzeroll J, Sleiman HF. Cyclometalated iridium(III) imidazole phenanthroline complexes as luminescent and electrochemiluminescent G-quadruplex DNA binders. Inorg Chem. 2015;54:6958–67.
Liang W, Zhuo Y, Xiong C, Zheng Y, Chai Y, Yuan R. Ultrasensitive cytosensor based on self-enhanced electrochemiluminescent ruthenium-silica composite nanoparticles for efficient drug screening with cell apoptosis monitoring. Anal Chem. 2015;87:12363–71.
Kadimisetty K, Malla S, Sardesai N P, Joshi AA, Faria RC, Lee NH, et al. Automated multiplexed ECL immunoarrays for cancer biomarker proteins. Anal Chem. 2015;87:4472–8.
Valenti G, Zangheri M, Sansaloni SE, Mirasoli M, Penicaud A, Roda A, et al. Transparent carbon nanotube network for efficient electrochemiluminescence devices. Chem Eur J. 2015;21:12640–5.
Yao H, Zhang Y, Xiao F, Xia Z, Rao J. Quantum dot/bioluminescence resonance energy transfer base highly sensitive detection of proteases. Angew Chem Int Ed. 2007;46:4346–9.
Shimidzu T, Iyoda T, Izaki K. Photoelectrochemical properties of bis(2,2′-bipyridlne) (4,4′-dlcarboxy-2,2′-bipyrldlne)ruthenlum(II) chloride. J Phys Chem. 1985;89:642–5.
Miao W, Choi JP, Bard AJ. Electrogenerated chemiluminescence 69: the tris(2,2‘-bipyridine)ruthenium(II), (Ru(bpy)32+)/Tri-n-propylamine (TPrA) system revisited-a new route involving TPrA·+cation radicals. J Am Chem Soc. 2002;124:14478–85.
Cho TJ, Zangmeister RA, Maccuspie RI, Patri AK, Hackley V. Newkome-type dendron-stabilized gold nanoparticles: synthesis, reactivity, and stability. Chem Mater. 2011;23:2665–76.
Bard AJ, Faulkner LR. Electrochemical methods, fundamentals and applications. 2nd ed. INC.: John Wiley & Sons; 2000.
Herne TM, Tarlov MJ. Characterization of DNA probes immobilized on gold surfaces. J Am Chem Soc. 1997;119:8916–20.
Adjémian J, Anne A, Cauet G, Demaille C. Cleavage-sensing redox peptide monolayers for the rapid measurement of the proteolytic activity of trypsin and α-thrombin enzymes. Langmuir. 2010;26:10347–56.
Wong T-S, Kwong DL-W, Sham JS-T, Wei WI, Kwong Y-L, Yuen AP-W. Clinicopathologic significance of plasma matrix metalloproteinase-2 and -9 levels in patients with undifferentiated nasopharyngeal carcinoma. Eur J Surg Oncol. 2004;30:560–4.
Miller C, Cuendet P, Graetzel M. Adsorbed ω-hydroxy thiol monolayers on gold electrodes: evidence for electron tunneling to redox species in solution. J Phys Chem. 1991;95:877–86.
Sun B, Qi H, Ma F, Gao Q, Zhang C, Miao W. Double covalent coupling method for the fabrication of highly sensitive and reusable electrogenerated chemiluminescence sensors. Anal Chem. 2010;82:5046–52.
Li Y, Qi H, Fang F, Zhang C. Ultrasensitive electrogenerated chemiluminescence detection of DNA hybridization using carbon-nanotubes loaded with tris(2,2’-bipyridyl) ruthenium derivative tags. Talanta. 2007;72:1704–9.
Chau Y, Pader RF, Dang NM, Langer R. Antitumor efficacy of a novel polymer–peptide–drug conjugate in human tumor xenograft models. Int J Cancer. 2006;118:1519–26.
Feng D, Zhang Y, Feng T, Shi W, Li X, Ma H. A graphene oxide–peptide fluorescence sensor tailor-made for simple and sensitive detection of matrix metalloproteinase 2. Chem Commun. 2011;47:10680–2.
Acknowledgments
We thank The National Science Foundation of China (Nos. 21522504, 21375084, 21275095, 21475082), the Natural Science Basic Research Plan in Shaanxi Province of China (No. 2014JQ2065, 2013SZS08-Z01, and 2013SZS08-P01), and the Fundamental Research Funds for the Central Universities (No. GK201505008).
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Published in the topical collection Analytical Electrochemiluminescence with guest editors Hua Cui, Francesco Paolucci, Neso Sojic, and Guobao Xu.
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Dang, Q., Gao, H., Li, Z. et al. Simple and sensitive electrogenerated chemiluminescence peptide-based biosensor for detection of matrix metalloproteinase 2 released from living cells. Anal Bioanal Chem 408, 7067–7075 (2016). https://doi.org/10.1007/s00216-016-9360-z
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DOI: https://doi.org/10.1007/s00216-016-9360-z