基于DNA折纸的酶级联催化用于低密度脂蛋白的电化学检测

引用本文: 余紫荆, 肖明书, 裴昊, 李丽. 基于DNA折纸的酶级联催化用于低密度脂蛋白的电化学检测[J]. 分析化学, 2022, 50(6): 850-858. doi: 10.19756/j.issn.0253-3820.221111 shu

Citation: YU Zi-Jing, XIAO Ming-Shu, PEI Hao, LI Li. DNA Origami-based Enzyme Cascade Catalysis for Electrochemical Detection of Low Density Lipoprotein[J]. Chinese Journal of Analytical Chemistry, 2022, 50(6): 850-858. doi: 10.19756/j.issn.0253-3820.221111 shu

基于DNA折纸的酶级联催化用于低密度脂蛋白的电化学检测

余紫荆 ^1, ,
肖明书 ^1, ,
裴昊 ^1, ,
李丽 ^{1,2,
,}

1.
华东师范大学化学与分子工程学院, 上海市绿色化学与化学过程重点实验室, 上海 200241;
2.
崇明生态研究院, 上海 202162

通讯作者: 李丽,E-mail:lli@chem.ecnu.edu.cn

基金项目:
国家自然科学基金项目（Nos.22174046，22104038）、上海市科学技术委员会（Nos.22ZR1419800，18490740500）和中国博士后科学基金项目（Nos.2020TQ0097，2021M701212）资助。

摘要: 很多疾病（如动脉粥样硬化）的发生与低密度脂蛋白（LDL）水平的上升密切相关，发展快速、灵敏的LDL分析方法对于相关疾病的早期诊断具有重要意义。本研究构建了一种基于DNA折纸的酶级联催化电化学传感器用于LDL的检测。基于核酸的可编程性和可寻址性，可将胆固醇氧化酶和辣根过氧化物酶特异性锚定在DNA折纸上，并通过控制酶间距（如20、40和80nm），实现了对LDL级联催化效率的优化。通过Au—S化学键，将负载酶的DNA折纸结构修饰在金电极表面，构建了酶级联催化电化学传感器。此传感器对LDL检测具有优异性能，在2~15μmol/L浓度范围内具有良好的线性关系，并且检出限低（1.8μmol/L （S/N=3）），选择性良好。将此传感器应用于健康人血清中LDL的检测，加标回收率在97.8%~103.6%之间，表明此传感器具有良好的重现性、稳定性和实用性。基于DNA折纸的酶级联催化电化学传感器为LDL的快速、灵敏检测提供了一种新方法，有望用于动脉粥样硬化的早期诊断。

关键词:

English

DNA Origami-based Enzyme Cascade Catalysis for Electrochemical Detection of Low Density Lipoprotein

YU Zi-Jing ^1, ,
XIAO Ming-Shu ^1, ,
PEI Hao ^1, ,
LI Li ^{1,2,
,}

1.
Shanghai Key Laboratory of Green Chemistry and Chemical Processes, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241, China;
2.
Institute of Eco-Chongming, Shanghai 202162, China

Corresponding author: LI Li, lli@chem.ecnu.edu.cn

Received Date: 01 March 2022
Revised Date: 21 March 2022
Fund Project:
Supported by the National Natural Science Foundation of China(Nos. 22074041, 22104038), the Science and Technology Commission of Shanghai Municipality(Nos. 22ZR1419800, 18490740500) and the China Postdoctoral Science Foundation(Nos. 2020TQ0097, 2021M701212).

Abstract: The development of many diseases(e.g., atherosclerosis) is closely associated with abnormal level of low density lipoprotein(LDL), and developing methods for rapid and sensitive detection of LDL is of great importance to early diagnosis of atherosclerosis. In this work, a novel electrochemical sensor based on DNA origami-based enzymatic cascade catalysis was reported for detection of LDL. Benefiting from the programmability and addressability of nucleic acids, two enzymes(i.e., cholesterol oxidase and horseradish peroxidase) were specifically assembled on DNA origami, and the cascade catalysis efficiency towards LDL was optimized by regulating their distance, including 20, 40 and 80 nm. Through Au-S chemical bond, DNA origami loading enzymes was modified on Au electrode to construct enzymatic cascade catalysis-based electrochemical sensor. The results demonstrated that this sensor showed excellent performance for LDL detection, with a linear response range of 2-15 μmol/L, a low limit of detection of 1.8 μmol/L(S/N=3), and good selectivity. The recoveries of LDL spiked in healthy human serum sample were 97.8%-103.6%, indicating good reproducibility and stability. This electrochemical sensor utilizing DNA origami-based enzymatic cascade catalysis provided a promising tool for rapid and sensitive detection of LDL, holding a great potential for early diagnosis of atherosclerosis.

Key words:

1. [1]
  LIBBY P. Nature, 2021, 592(7855):524-533.LIBBY P. Nature, 2021, 592(7855):524-533.
2. [2]
  GLASS C K, WITZTUM J L. Cell, 2001, 104(4):503-516.GLASS C K, WITZTUM J L. Cell, 2001, 104(4):503-516.
3. [3]
  FERENCE B A, GINSBERG H N, GRAHAM I, RAY K K, PACKARD C J, BRUCKERT E, HEGELE R A, KRAUSS R M, RAAL F J, SCHUNKERT H. Eur. Heart J., 2017, 38(32):2459-2472.FERENCE B A, GINSBERG H N, GRAHAM I, RAY K K, PACKARD C J, BRUCKERT E, HEGELE R A, KRAUSS R M, RAAL F J, SCHUNKERT H. Eur. Heart J., 2017, 38(32):2459-2472.
4. [4]
  BOREN J, CHAPMAN M J, KRAUSS R M, PACKARD C J, BENTZON J F, BINDER C J, DAEMEN M J, DEMER L L, HEGELE R A, NICHOLLS S J. Eur. Heart J., 2020, 41(24):2313-2330.BOREN J, CHAPMAN M J, KRAUSS R M, PACKARD C J, BENTZON J F, BINDER C J, DAEMEN M J, DEMER L L, HEGELE R A, NICHOLLS S J. Eur. Heart J., 2020, 41(24):2313-2330.
5. [5]
  MCMAHON M, GROSSMAN J, FITZGERALD J, DAHLIN-LEE E, WAKKACE D J, THONG B Y, BADSHA H, KALUNIAN K, CHARLES C, NAVAB M, FOGELMAN A M, HAHN B H. Arthritis Rheum., 2006, 54(8):2541-2549.MCMAHON M, GROSSMAN J, FITZGERALD J, DAHLIN-LEE E, WAKKACE D J, THONG B Y, BADSHA H, KALUNIAN K, CHARLES C, NAVAB M, FOGELMAN A M, HAHN B H. Arthritis Rheum., 2006, 54(8):2541-2549.
6. [6]
  TRPKOVIC A, RESANOVIC I, STANIMIROVIC J, RADAK D, MOUSA S A, CENIC-MILOSEVIC D, JEVREMOVIC D, ISENOVIC E R. Crit. Rev. Clin. Lab. Sci., 2015, 52(2):70-85.TRPKOVIC A, RESANOVIC I, STANIMIROVIC J, RADAK D, MOUSA S A, CENIC-MILOSEVIC D, JEVREMOVIC D, ISENOVIC E R. Crit. Rev. Clin. Lab. Sci., 2015, 52(2):70-85.
7. [7]
  DONG J, GUO H J, YANG R Y, LI H X, WANG S, ZHANG J T, ZHOU W Y, CHEN W X. Clin. Chim. Acta, 2012, 413(13-14):1071-1076.DONG J, GUO H J, YANG R Y, LI H X, WANG S, ZHANG J T, ZHOU W Y, CHEN W X. Clin. Chim. Acta, 2012, 413(13-14):1071-1076.
8. [8]
  TAN X W, TAKENAKA F, TAKEKAWA H, MATSUURA E. Heliyon, 2020, 6(6):e04114TAN X W, TAKENAKA F, TAKEKAWA H, MATSUURA E. Heliyon, 2020, 6(6):e04114
9. [9]
  BERNEIS K, LA BELLE M, BLANCHE P J, KRAUSS R M. J. Lipid Res., 2002, 43(7):1155-1159.BERNEIS K, LA BELLE M, BLANCHE P J, KRAUSS R M. J. Lipid Res., 2002, 43(7):1155-1159.
10. [10]
  JIE G F, LIU B, PAN H C, ZHU J J, CHEN H Y. Anal. Chem., 2007, 79(15):5574-5581.JIE G F, LIU B, PAN H C, ZHU J J, CHEN H Y. Anal. Chem., 2007, 79(15):5574-5581.
11. [11]
  ALI M A, SOLANKI P R, SRIVASTAVA S, SINGH S, AGRAWAL V V, JOHN R, MALHOTRA B D. ACS Appl.Mater. Interfaces, 2015, 7(10):5837-5846.ALI M A, SOLANKI P R, SRIVASTAVA S, SINGH S, AGRAWAL V V, JOHN R, MALHOTRA B D. ACS Appl.Mater. Interfaces, 2015, 7(10):5837-5846.
12. [12]
  WANG X, PEREIRA J H, TSUTAKAWA S, FANG X, ADAMS P D, MUKHOPADHYAY A, LEE T S. Metab.Eng., 2021, 64:41-51.WANG X, PEREIRA J H, TSUTAKAWA S, FANG X, ADAMS P D, MUKHOPADHYAY A, LEE T S. Metab.Eng., 2021, 64:41-51.
13. [13]
  FU Y M, ZENG D D, CHAO J, JIN Y Q, ZHANG Z, LIU Hu J, LI D, MA H W, HUANG Q, GOTHELF K V. J.Am. Chem. Soc., 2013, 135(2):696-702.FU Y M, ZENG D D, CHAO J, JIN Y Q, ZHANG Z, LIU Hu J, LI D, MA H W, HUANG Q, GOTHELF K V. J.Am. Chem. Soc., 2013, 135(2):696-702.
14. [14]
  XIAO M S, LAI W, MAN T T, CHANG B B, LI L, CHANDRASEKARAN A R, PEI H. Chem. Rev., 2019, 119(22):11631-11717.XIAO M S, LAI W, MAN T T, CHANG B B, LI L, CHANDRASEKARAN A R, PEI H. Chem. Rev., 2019, 119(22):11631-11717.
15. [15]
  YU Li-Xing, ZHAI Rui, GONG Xiao-Yun, XIE Jie, HUANG Ze-Jian, LIU Mei-Ying, JIANG You, DAI Xin-Hua, FANG Xiang, YU Xiao-Ping. Chin. J. Anal. Chem., 2019, 47(11):1742-1750.余利星, 翟睿, 龚晓云, 谢洁, 黄泽建, 刘梅英, 江游, 戴新华, 方向, 俞晓平.分析化学, 2019, 47(11):1742-1750.
16. [16]
  CAO M, SUN Y, XIAO M, LI L, LIU X, JIN H, PEI H. Chem. Res. Chin. Univ., 2020, 36(2):254-260.CAO M, SUN Y, XIAO M, LI L, LIU X, JIN H, PEI H. Chem. Res. Chin. Univ., 2020, 36(2):254-260.
17. [17]
  JI W, LI X, XIAO M, SUN Y, LAI W, ZHANG H, PEI H, LI L. Chem.-Eur. J., 2021. 27(34):8745-8752.JI W, LI X, XIAO M, SUN Y, LAI W, ZHANG H, PEI H, LI L. Chem.-Eur. J., 2021. 27(34):8745-8752.
18. [18]
  HAN D, PAL S, NANGREAVE J, DENG Z, LIU Y, YAN H. Science, 2011, 332(6027):342-346.HAN D, PAL S, NANGREAVE J, DENG Z, LIU Y, YAN H. Science, 2011, 332(6027):342-346.
19. [19]
  HONG F, ZHANG F, LIU Y, YAN H. Chem. Rev., 2017, 117(20):12584-12640.HONG F, ZHANG F, LIU Y, YAN H. Chem. Rev., 2017, 117(20):12584-12640.
20. [20]
  MALLIK L, DHAKAL S, NICHOLS J, MAHONEY J, DOSEY A M, JIANG S X, SUNAHARA R K, SKINIOTIS G, WALTER N G. ACS Nano, 2015, 9(7):7133-7141.MALLIK L, DHAKAL S, NICHOLS J, MAHONEY J, DOSEY A M, JIANG S X, SUNAHARA R K, SKINIOTIS G, WALTER N G. ACS Nano, 2015, 9(7):7133-7141.
21. [21]
  PEI H, SHA R J, WANG X W, ZHENG M, FAN C H, CANARY J W, SEEMAN N C. J. Am. Chem. Soc., 2019, 141(30):11923-11928.PEI H, SHA R J, WANG X W, ZHENG M, FAN C H, CANARY J W, SEEMAN N C. J. Am. Chem. Soc., 2019, 141(30):11923-11928.
22. [22]
  XIAO M S, LAI W, WANG F, LI L, FAN C H, PEI H. J. Am. Chem. Soc., 2019, 141(51):20354-20364.XIAO M S, LAI W, WANG F, LI L, FAN C H, PEI H. J. Am. Chem. Soc., 2019, 141(51):20354-20364.
23. [23]
  ZHANG Tong, TAO Qing, BIAN Xiao-Jun, CHEN Qian, YAN Juan. Chin. J. Anal. Chem., 2021, 49(3):377-386.张彤, 陶晴, 卞晓军, 陈谦, 颜娟.分析化学, 2021, 49(3):377-386.
24. [24]
  VENEZIANO R, MOYER T J, STONE M B, WAMHOFF E C, READ B J, MUKHERJEE S, SHEPHERD T R, DAS J, SCHIEF W R, IRVINE D J, BATHE M. Nat. Nanotehnol., 2020, 15(8):716-723.VENEZIANO R, MOYER T J, STONE M B, WAMHOFF E C, READ B J, MUKHERJEE S, SHEPHERD T R, DAS J, SCHIEF W R, IRVINE D J, BATHE M. Nat. Nanotehnol., 2020, 15(8):716-723.
25. [25]
  HONG F, ZHANG F, LIU Y, YAN H. Chem. Rev., 2017, 117(20):12584-12640.HONG F, ZHANG F, LIU Y, YAN H. Chem. Rev., 2017, 117(20):12584-12640.
26. [26]
  PUCHKOVA A, VIETZ C, PIBIRI E, WUNSCH B, SANZ-PAZ M, ACUNA G P, TINNEFELD P. Nano Lett., 2015, 15(12):8354-8359.PUCHKOVA A, VIETZ C, PIBIRI E, WUNSCH B, SANZ-PAZ M, ACUNA G P, TINNEFELD P. Nano Lett., 2015, 15(12):8354-8359.
27. [27]
  OCHMANN S E, VIETZ C, TROFYMCHUK K, ACUNA G P, LALKENS B, TINNEFELD P. Anal. Chem. 2017, 89(23):13000-13007.OCHMANN S E, VIETZ C, TROFYMCHUK K, ACUNA G P, LALKENS B, TINNEFELD P. Anal. Chem. 2017, 89(23):13000-13007.
28. [28]
  ZHAO D, KONG Y, ZHAO S, XING H. Top. Curr. Chem., 2020, 378:41.ZHAO D, KONG Y, ZHAO S, XING H. Top. Curr. Chem., 2020, 378:41.
29. [29]
  NIEMEYER C M. Angew. Chem., Int. Ed., 2010, 49(7):1200-1216.NIEMEYER C M. Angew. Chem., Int. Ed., 2010, 49(7):1200-1216.
30. [30]
  FU J, YANG Y R, DHAKAL S, ZHAO Z, LIU M, ZHANG T, WALTER N G, YAN H. Nat. Protoc., 2016, 11(11):2243-2273.FU J, YANG Y R, DHAKAL S, ZHAO Z, LIU M, ZHANG T, WALTER N G, YAN H. Nat. Protoc., 2016, 11(11):2243-2273.
31. [31]
  FU J, LIU M, LIU Y, WOOBURY N W, YAO H. J. Am. Chem. Soc., 2012, 134(12):5516-5519.FU J, LIU M, LIU Y, WOOBURY N W, YAO H. J. Am. Chem. Soc., 2012, 134(12):5516-5519.
32. [32]
  KAHN J S, XIONG Y, HUANG J, GANG O. JACS Au, 2022, 2(2):357-366.KAHN J S, XIONG Y, HUANG J, GANG O. JACS Au, 2022, 2(2):357-366.
33. [33]
  SONG P, SHEN J, YE D, DONG B, WANG F, PEI H, WANG J, SHI J, WANG L, XUE W, HUANG Y, HUANG G, ZUO X, FAN C. Nat. Commun., 2020, 11:838.SONG P, SHEN J, YE D, DONG B, WANG F, PEI H, WANG J, SHI J, WANG L, XUE W, HUANG Y, HUANG G, ZUO X, FAN C. Nat. Commun., 2020, 11:838.
34. [34]
  KE Y G, LINDSAY S, CHANG Y, LIU Y, YAN H. Science, 2008, 319(5860):180-183.KE Y G, LINDSAY S, CHANG Y, LIU Y, YAN H. Science, 2008, 319(5860):180-183.
35. [35]
  GE Z, FU J, LIU M, JIANG S, ANDREONI A, ZUO X, LIU Y, YAN H, FAN C. ACS Appl. Mater. Interfaces, 2018, 11(15):13881-13887.GE Z, FU J, LIU M, JIANG S, ANDREONI A, ZUO X, LIU Y, YAN H, FAN C. ACS Appl. Mater. Interfaces, 2018, 11(15):13881-13887.
36. [36]
  ZHANG J, SONG S P, WANG L H, PAN D, FAN C H. Nat. Protoc., 2007, 2(11):2888-2895.ZHANG J, SONG S P, WANG L H, PAN D, FAN C H. Nat. Protoc., 2007, 2(11):2888-2895.
37. [37]
  ALI M A, KAMIL R K, SRIVASTAVA S, AGRAWAL V V, JOHN R, MALHOTRA B D. Langmuir, 2014, 30(14):4192-4201.ALI M A, KAMIL R K, SRIVASTAVA S, AGRAWAL V V, JOHN R, MALHOTRA B D. Langmuir, 2014, 30(14):4192-4201.
38. [38]
  ALI M A, SRIVASTAVA S, PANDEY M K, AGRAWAL V V, JOHN R, MALHOTRA B D. Anal. Chem., 2014, 86(3):1710-1718.ALI M A, SRIVASTAVA S, PANDEY M K, AGRAWAL V V, JOHN R, MALHOTRA B D. Anal. Chem., 2014, 86(3):1710-1718.
39. [39]
  LIU Y, YU D S, ZENG C, MIAO Z C, DAI L M. Langmuir, 2010, 26(9):6158-6160LIU Y, YU D S, ZENG C, MIAO Z C, DAI L M. Langmuir, 2010, 26(9):6158-6160
40. [40]
  YARA A, MANEL V. Electrochim. Acta, 2017, 229:458-466.YARA A, MANEL V. Electrochim. Acta, 2017, 229:458-466.
41. [41]
  ALI M A, SIINGH C, MONDAL K, SRIVASTAVA S, SHARMA A, MALHOTRA B D. ACS Appl. Mater.Interfaces, 2016, 8(12):7646-7656.ALI M A, SIINGH C, MONDAL K, SRIVASTAVA S, SHARMA A, MALHOTRA B D. ACS Appl. Mater.Interfaces, 2016, 8(12):7646-7656.

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于DNA折纸的酶级联催化用于低密度脂蛋白的电化学检测

通讯作者: 李丽,E-mail:lli@chem.ecnu.edu.cn

English

DNA Origami-based Enzyme Cascade Catalysis for Electrochemical Detection of Low Density Lipoprotein

Corresponding author: LI Li, lli@chem.ecnu.edu.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于DNA折纸的酶级联催化用于低密度脂蛋白的电化学检测

通讯作者: 李丽,E-mail:lli@chem.ecnu.edu.cn

English

DNA Origami-based Enzyme Cascade Catalysis for Electrochemical Detection of Low Density Lipoprotein

Corresponding author: LI Li, lli@chem.ecnu.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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