New principle for the direct real-time monitoring of
interaction of cholinesterase and its inhibitors

J 2003

New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors

MAKOWER, Alexander, Jan HALÁMEK, Petr SKLÁDAL, Franz KERNCHENC, Frieder SCHELLER et. al.

Basic information

Original name

New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors

Authors

MAKOWER, Alexander (276 Germany), Jan HALÁMEK (203 Czech Republic), Petr SKLÁDAL (203 Czech Republic, guarantor), Franz KERNCHENC (276 Germany) and Frieder SCHELLER (276 Germany)

Edition

Biosensors & Bioelectronics, Oxford, Elsevier Advanced Technology, 2003, 0956-5663

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 2.947

RIV identification code

RIV/00216224:14310/03:00008862

Organization unit

Faculty of Science

Keywords in English

cholinesterase; inhibitor; piezoelectric; biosensor; real-time

Abstract

V originále

A new method for the sensitive detection of cholinesterase inhibitors based on real-time monitoring using a piezoelectric biosensor. The cholinesterase inhibitor paraoxon was immobilized on the sensing surface via a chelate complex as the recognition element. At first, the conjugate of N?mercaptoundecanoic acid (MUA) with Ná, Ná-bis (carboxymethyl)-L-Lysine (NTA-Lys) was chemisorbed to form a self-assembled monolayer on the surface of the gold electrode of the piezosensor. In the next step, paraoxon-spacer-hexahistidine conjugate was linked to the MUA-Lys-NTA layer via the chelate complex with Ni2+. The paraoxon-modified surface thus obtained was applied for the binding of human butyrylcholinesterase. Regeneration of the sensing surface was achieved by splitting the chelate complex with EDTA and depositing a fresh layer of Ni2+ followed by addition of the paraoxon-spacer-hexahistidine. In the presence of free inhibitors like diisopropylfluorphosphate (DFP), binding of BChE to the surface-bound paraoxon was decreased. In this way, a competitive affinity assay for organophosphorus compounds was developed. The limit of detection for DFP as a model compound was 10 nmol/l (approx. 2 mg/l). This new concept seems suitable for constructing biosensors for the group-specific detection of cholinesterase-inhibiting substances like insecticides in the field.

Links

OC 518.30, research and development project

Name: Sledování interakcí biomakromolekul s biovrstvami v různých konformačních stavech pomocí piezoelektrických biosensorů

Investor: Ministry of Education, Youth and Sports of the CR, Characterization of interactions of biomacromolecules with biolayers in different conformation states using piezoelectric biosensors

Citovat

MAKOWER, Alexander, Jan HALÁMEK, Petr SKLÁDAL, Franz KERNCHENC and Frieder SCHELLER. New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors. Biosensors & Bioelectronics. Oxford: Elsevier Advanced Technology, 2003, vol. 18, No 11, p. 1329-1337. ISSN 0956-5663.

@article{488275,
   author = {Makower, Alexander and Halámek, Jan and Skládal, Petr and Kernchenc, Franz and Scheller, Frieder},
   article_location = {Oxford},
   article_number = {11},
   keywords = {cholinesterase; inhibitor; piezoelectric; biosensor; real-time},
   language = {eng},
   issn = {0956-5663},
   journal = {Biosensors & Bioelectronics},
   title = {New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors},
   volume = {18},
   year = {2003}
}

TY  - JOUR
ID  - 488275
AU  - Makower, Alexander - Halámek, Jan - Skládal, Petr - Kernchenc, Franz - Scheller, Frieder
PY  - 2003
TI  - New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors
JF  - Biosensors & Bioelectronics
VL  - 18
IS  - 11
SP  - 1329-1337
EP  - 1329-1337
PB  - Elsevier Advanced Technology
SN  - 09565663
KW  - cholinesterase
KW  - inhibitor
KW  - piezoelectric
KW  - biosensor
KW  - real-time
N2  - A new method for the sensitive detection of cholinesterase inhibitors based on real-time monitoring using a piezoelectric biosensor. The cholinesterase inhibitor paraoxon was immobilized on the sensing surface via a chelate complex as the recognition element. At first, the conjugate of N?mercaptoundecanoic acid (MUA) with Ná, Ná-bis (carboxymethyl)-L-Lysine (NTA-Lys) was chemisorbed to form a self-assembled monolayer on the surface of the gold electrode of the piezosensor. In the next step, paraoxon-spacer-hexahistidine conjugate was linked to the MUA-Lys-NTA layer via the chelate complex with Ni2+. The paraoxon-modified surface thus obtained was applied for the binding of human butyrylcholinesterase. Regeneration of the sensing surface was achieved by splitting the chelate complex with EDTA and depositing a fresh layer of Ni2+ followed by addition of the paraoxon-spacer-hexahistidine. In the presence of free inhibitors like diisopropylfluorphosphate (DFP), binding of BChE to the surface-bound paraoxon was decreased. In this way, a competitive affinity assay for organophosphorus compounds was developed. The limit of detection for DFP as a model compound was 10 nmol/l (approx. 2 mg/l). This new concept seems suitable for constructing biosensors for the group-specific detection of cholinesterase-inhibiting substances like insecticides in the field.
ER  -

MAKOWER, Alexander, Jan HALÁMEK, Petr SKLÁDAL, Franz KERNCHENC and Frieder SCHELLER. New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors. \textit{Biosensors \&{} Bioelectronics}. Oxford: Elsevier Advanced Technology, 2003, vol.~18, No~11, p.~1329-1337. ISSN~0956-5663.

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