In situ grown DNA nanotail-templated silver nanoclusters enabling label-free electrochemical sensing of terminal deoxynucleotidyl transferase activity

被引：43

作者：

Hu, Yufang ^{[1
]}

Zhang, Qingqing ^{[1
]}

Guo, Zhiyong ^{[1
]}

Wang, Sui ^{[1
]}

Du, Chunnuan ^{[1
]}

Zhai, Chunyang ^{[1
]}

机构：

[1] Ningbo Univ, Fac Mat Sci & Chem Engn, Ningbo 315211, Zhejiang, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2017年 / 98卷

基金：

中国国家自然科学基金;

关键词：

Terminal deoxynucleotidyl transferase; Silver nanoclusters; Graphene oxide; Enzyme activity; Inhibitor; ENZYME-ACTIVITY; GRAPHENE OXIDE; METAL NANOCLUSTERS; AMPLIFICATION; NANOPARTICLES; LEUKEMIA; ASSAY; DIAGNOSIS; MICRORNA; SENSOR;

D O I：

10.1016/j.bios.2017.06.017

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

A novel label-free electrochemical strategy was established based on the unique electro-catalytic activity of graphene oxide (GO)-supported terminal deoxynucleotidyl transferase (TM-generated C-rich DNA nanotail-templated silver nanoclusters (DNA-AgNCs). TdT can catalyze the deoxycytidine triphosphate (dCTP) to the 3'-OH terminus of single-stranded DNA (ssDNA) with no template; then, in the presence of Ag(I), TdT-generated C-rich DNA sequence was employed for the synthetic template of AgNCs because of the formed complexes of nitrogen atoms of cytosine based with silver atoms. We proved that in situ grown DNA nanotail-templated AgNCs can be adsorbed on GO-modified electrode and possess high electro-catalytic activity to H2O2 reduction, presenting a good electrochemical indicator for signal readout. Under optimal conditions, the proposed biosensor could be employed for quantitatively monitoring TdT activity and within a dynamic range from 0.4 to 90 U/mL and a low limit of detection is 0.08 U/mL. With high sensitivity and excellent selectivity, this strategy offers a facile, convenient and specific electrochemical method for TdT activity detection and its relevant inhibitors screening. It holds a promising potential in the practical application of TdT-based biochemical research, disease diagnosis and drug discovery.

引用

页码：91 / 99

页数：9

共 39 条

[1] Increased junctional diversity in fetal B cells results in a loss of protective anti-phosphorylcholine antibodies in adult mice [J].

Benedict, CL ;

Kearney, JF .

IMMUNITY, 1999, 10 (05) :607-617

[2]

Benedict CL, 2000, IMMUNOL REV, V175, P150, DOI 10.1111/j.1600-065X.2000.imr017518.x

[3] CHEMICALLY DEFINED TEMPLATES + INITIATORS FOR DEOXYPOLYNUCLEOTIDE SYNTHESIS [J].

BOLLUM, FJ .

SCIENCE, 1964, 144 (361) :560-&

[4]

BOLLUM FJ, 1960, J BIOL CHEM, V235, P2399

[5] Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles [J].

Elghanian, R ;

Storhoff, JJ ;

Mucic, RC ;

Letsinger, RL ;

Mirkin, CA .

SCIENCE, 1997, 277 (5329) :1078-1081

[6] A peptide with a cysteine terminus: probe for label-free fluorescent detection of thrombin activity [J].

Feng, Jingjing ;

Zhuo, Caixia ;

Ma, Xuejuan ;

Li, Shuangqin ;

Zhang, Yaodong .

ANALYST, 2016, 141 (14) :4481-4487

[7] DEMONSTRATION OF TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE IN THYMOCYTES BY IMMUNOFLUORESCENCE [J].

GOLDSCHNEIDER, I ;

GREGOIRE, KE ;

BARTON, RW ;

BOLLUM, FJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (02) :734-738

[8]

Gopalakrishnan AM., 2003, NATURE, V421, P431

[9] ACUTE LYMPHOBLASTIC-LEUKEMIA ASSOCIATED ANTIGEN .3. ALTERATIONS IN EXPRESSION DURING TREATMENT AND IN RELAPSE [J].

GREAVES, M ;

PAXTON, A ;

JANOSSY, G ;

PAIN, C ;

JOHNSON, S ;

LISTER, TA .

LEUKEMIA RESEARCH, 1980, 4 (01) :1-14

[10]

HOFFBRAND AV, 1977, LANCET, V2, P520

← 1 2 3 4 →