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High efficiency acetylcholinesterase immobilization on DNA aptamer modified surfaces

Title: High efficiency acetylcholinesterase immobilization on DNA aptamer modified surfaces
Authors: Chumphukam, O
Le, TT
Cass, AEG
Item Type: Journal Article
Abstract: 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 (Kd = 157 ± 42 pM). Characterization of the aptamer showed its binding is not affected by low ionic strength (~20 mM), however significant reduction in affinity occurred at high ionic strength (~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.
Issue Date: 21-Apr-2014
Date of Acceptance: 11-Apr-2014
URI: http://hdl.handle.net/10044/1/68248
DOI: https://dx.doi.org/10.3390/molecules19044986
ISSN: 1420-3049
Publisher: MDPI AG
Start Page: 4986
End Page: 4996
Journal / Book Title: Molecules
Volume: 19
Issue: 4
Copyright Statement: © 2014 The Author(s). This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0 - https://creativecommons.org/licenses/by/3.0/).
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Engineering & Physical Science Research Council (EPSRC)
Funder's Grant Number: BB/I001824/1
EP/K039946/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemistry & Molecular Biology
Chemistry, Multidisciplinary
Chemistry
aptamers
acetylcholinesterase
immobilization
SINGLE-STRANDED-DNA
PROTEIN-PURIFICATION
MONOVALENT CATIONS
HUMAN THROMBIN
AFFINITY TAGS
MICROARRAYS
RNA
PERFORMANCE
ANTIBODIES
LIGANDS
Acetylcholinesterase
Animals
Aptamers, Nucleotide
Base Sequence
Electrophorus
Enzymes, Immobilized
Fish Proteins
Kinetics
Molecular Sequence Data
Nucleic Acid Conformation
Osmolar Concentration
Protein Binding
SELEX Aptamer Technique
Streptavidin
0305 Organic Chemistry
Organic Chemistry
Publication Status: Published
Online Publication Date: 2014-04-21
Appears in Collections:Chemistry
Biological and Biophysical Chemistry
Faculty of Natural Sciences



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