基于表面增强拉曼光谱的酪氨酸酶可抛式传感器研究
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摘要
研制了一种可抛式表面增强拉曼光谱(SERS)传感器用于检测血清中酪氨酸酶(TYR)活性。首先制备了功能化银纳米颗粒浆料,采用丝网印刷技术制备SERS传感器。修饰在传感器上的4-巯基邻苯二酚可被酪氨酸酶催化氧化为4-巯基邻苯二醌,引起传感器SERS光谱的显著变化,从而实现对酪氨酸酶的检测。此传感器对酪氨酸酶响应快速,在优化条件下5 min内即可完成检测;特征SERS信号强度与酪氨酸酶浓度在1～100 U/mL范围内呈良好的线性关系,检出限为0.28 U/mL(3σ)。由于检测反应的特异性和SERS技术可提供指纹信息的综合优势,此传感器有较好的抗干扰性能。血清样品中的加标回收率为88.1%～104.6%,相对标准偏差(RSD)小于4%,可准确反映苯甲酸的含量及其对酪氨酸酶的抑制能力。结果表明,所研制的可抛式SERS传感器可一次性使用,适合生物样品中酪氨酸酶的快速分析,以及酪氨酸酶抑制剂的快速筛选和检测。
A novel disposable surface-enhanced Raman scattering(SERS) sensor based on enzyme-catalyzed oxidation was developed for detecting tyrosinase(TYR) activity. The SERS sensor was fabricated by assembling the synthesized 4-thiol-catechol onto the surface of silver nanoparticles(AgNPs) which were screen printed on supporting material, and then the detection of TYR activity was accomplished according to SERS spectral changes of the SERS senor caused by the reaction between 4-thiol-catechol and TYR. The results showed that the SERS sensor had fast response to TYR(less than 5 min), and there was a linear relationship between the characteristic SERS signal and TYR concentration from 1 U/mL to 100 U/mL with limit of detection(LOD) of 0.28 U/mL(3σ). Moreover, resulting from the fingerprinting advantage of SERS technique and the specific property of the employed sensing-reaction, the developed SERS sensor had good selectivity towards TYR, which could be used for detection and evaluation of TYR and TYR inhibitor in serum.
A novel disposable surface-enhanced Raman scattering(SERS) sensor based on enzyme-catalyzed oxidation was developed for detecting tyrosinase(TYR) activity. The SERS sensor was fabricated by assembling the synthesized 4-thiol-catechol onto the surface of silver nanoparticles(AgNPs) which were screen printed on supporting material, and then the detection of TYR activity was accomplished according to SERS spectral changes of the SERS senor caused by the reaction between 4-thiol-catechol and TYR. The results showed that the SERS sensor had fast response to TYR(less than 5 min), and there was a linear relationship between the characteristic SERS signal and TYR concentration from 1 U/mL to 100 U/mL with limit of detection(LOD) of 0.28 U/mL(3σ). Moreover, resulting from the fingerprinting advantage of SERS technique and the specific property of the employed sensing-reaction, the developed SERS sensor had good selectivity towards TYR, which could be used for detection and evaluation of TYR and TYR inhibitor in serum.
引文
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2 Yang X M, Luo Y W, Zhuo Y, Feng Y J, Zhu S S. Anal. Chim. Acta, 2014, 840: 87-92
3 Baharav E, Merimski O, Shoenfeld Y, Zigelman R, Gilbrud B, Yecheskel G, Youinou P, Fishman P. Clin. Exp. Immunol., 1996, 105(1): 84-88
4 Ando H, Kondoh H, Ichihashi M, Hearing V J. J. Invest. Dermatol., 2007, 127(4): 751-761
5 Tessari I, Bisaglia M, Valle F, Samorì B, Bergantino E, Mammi S, Bubacco L.J. Biol. Chem., 2008, 283(24): 16808-16817
6 Angeletti C, Khomitch V, Halaban R, Rimm D L. Diagn. Cytopathol. , 2004, 31(1): 33-37
7 Zhu X L, Hu J, Zhao Z H, Sun M J, Chi X Q, Wang X M, Gao J H. Small, 2015, 11(7): 862-870
8 Liu B W, Huang P C, Li J F, Wu F Y. Sens. Actuators B, 2017, 251: 836-841
9 Li D, Gill R, Freeman R, Willner I. Chem. Commun., 2006, 1(48): 5027-5029
10 Wu X F, Li L H, Shi W, Gong Q Y, Ma H M. Angew. Chem. Int. Edit., 2016, 55(47): 14728-14732
11 Qu Z Y, Na W D, Liu X T, Liu H, Su X G. Anal. Chim. Acta, 2018, 997: 52-59
12 Li H H, Liu W, Zhang F Y, Zhu X Y, Huang L Q, Zhang H X. Anal. Chem., 2018, 90(1): 855-858
13 Ma X G, Gao W Y, Halawa M I, Lan Y X, Li J P, Xu G B. Sens. Actuators B, 2019, 280: 41-45
14 Hoebe R A, Van Oven C H, Gadella T W J, Dhonukshe P B, Van Noorden C J F, Manders E M M. Nat. Biotechnol., 2007, 25(2): 249-253
15 Luo L, Chen Y H, Zhang L X, Li Y R, Li H L, Zhang H Q, Tian Y. Microchim. Acta, 2017, 184(2): 595-601
16 Li Z P, Wang Y F, Zhang X, Zeng C C, Hu L M, Liang X J. Sens. Actuators B, 2017, 242: 189-194
17 Sabatt G, Keir R, Lawlor M, Black M, Graham D, Smith W E. Anal. Chem., 2008, 80(7): 2351-2356
18 Yang L B, Li P, Liu H L, Tang X H, Liu J H. Chem. Soc. Rev., 2015, 44(10): 2837-2848
19 Braun G, Lee S J, Dante M, Nguyen T Q, Moskovits M, Reich N. J. Am. Chem. Soc., 2007, 129(20): 6378-6379
20 Li P, Ma B B, Yang L B, Liu J H. Chem. Commun., 2015, 51(57): 11394-11397
21 Miquel A O, Rosa P, Sebastian W H, Angel M. Sens. Actuators B, 2017, 252: 657-662
22 Wang W K, Zhang L M, Li L, Tian Y. Anal. Chem., 2016, 88(19): 9518-9523
23 Li D W, Qu L L, Hu K, Long Y T, Tian H. Angew. Chem. Int. Edit., 2015, 54(43): 12758-12761
24 Joseph A M, Nagendra B, Gowd E B, Surendran K P. ACS Omega, 2016, 1(6): 1220-1228
25 Qu L L, Li D W, Xue J Q, Zhai W L, Fossey J S, Long Y T. Lab Chip, 2012, 12(5): 876-881
26 Zhao L J, Qian R C, Ma W, Tian H, Long Y T. Anal. Chem., 2016, 88(17): 8375-8379
27 Lee P C, Meisel D. J. Phys. Chem., 1982, 86(17): 3391-3395
28 Trefry J C, Monahan J L, Weaver K M, Meyerhoefer A J, Markopolous M M, Arnold Z S, Wooley D P, Pavel I E. J. Am. Chem. Soc., 2010, 132(32): 10970-10972
29 Liu Z P, Liu S S. Anal. Bioanal. Chem., 2018, 410(17): 4145-4152
30 Sidhu J S, Singh N. J. Mater. Chem. B, 2018, 6(24): 4139-4145
31 Wu X F, Li X H, Li H Y, Shi W, Ma H M. Chem. Commun., 2017, 53(16): 2443-2446
32 Ao H, Qian Z S, Zhu Y Y, Zhao M Z, Tang C, Huang Y Y, Feng H, Wang A J. Biosens. Bioelectron., 2016, 86: 542-547
33 Wang C C, Yan S Y, Huang R, Feng S, Fu B S, Weng X C, Zhou X. Analyst, 2013, 138(10): 2825-2828
34 Zhao J H, Bao X F, Wang S, Lu S S, Sun J, Yang X R. Anal.Chem., 2017, 89(19): 10529-10536