High Efficiency Acetylcholinesterase Immobilization on DNA Aptamer Modified Surfaces

被引：13

作者：

Chumphukam, Orada ^{[1
]}

Le, Thao T. ^{[1
]}

Cass, Anthony E. G. ^{[1
]}

机构：

[1] Univ London Imperial Coll Sci Technol & Med, Dept Chem, London SW7 2AZ, England

来源：

MOLECULES | 2014年 / 19卷 / 04期

基金：

英国生物技术与生命科学研究理事会; 英国工程与自然科学研究理事会;

关键词：

aptamers; acetylcholinesterase; immobilization; SINGLE-STRANDED-DNA; PROTEIN-PURIFICATION; MONOVALENT CATIONS; HUMAN THROMBIN; AFFINITY TAGS; MICROARRAYS; RNA; PERFORMANCE; ANTIBODIES; LIGANDS;

D O I：

10.3390/molecules19044986

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report here the in vitro selection of DNA aptamers for electric eel acetylcholinesterase (AChE). One selected aptamer sequence (R15/19) has a high affinity towards the enzyme (K-d = 157 +/- 42 pM). Characterization of the aptamer showed its binding is not affected by low ionic strength (similar to 20 mM), however significant reduction in affinity occurred at high ionic strength (similar to 1.2 M). In addition, this aptamer does not inhibit the catalytic activity of AChE that we exploit through immobilization of the DNA on a streptavidin-coated surface. Subsequent immobilization of AChE by the aptamer results in a 4-fold higher catalytic activity when compared to adsorption directly on to plastic.

引用

页码：4986 / 4996

页数：11

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