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.

Základní údaje

Originální název

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

Autoři

MAKOWER, Alexander; Jan HALÁMEK; Petr SKLÁDAL; Franz KERNCHENC a Frieder SCHELLER

Vydání

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

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 2.947

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/03:00008862

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

cholinesterase; inhibitor; piezoelectric; biosensor; real-time

Štítky

biosensor, Cholinesterase, inhibitor, piezoelectric, real-time

Změněno: 5. 9. 2003 14:11, prof. RNDr. Petr Skládal, CSc.

Anotace

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.

Návaznosti

OC 518.30, projekt VaV

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

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Sledování interakcí biomakromolekul s biovrstvami v různých konformačních stavech pomocí piezoelektrických biosensorů

Citovat

MAKOWER, Alexander; Jan HALÁMEK; Petr SKLÁDAL; Franz KERNCHENC a Frieder SCHELLER. New principle for the direct real-time monitoring of interaction of cholinesterase and its inhibitors. Biosensors & Bioelectronics. Oxford: Elsevier Advanced Technology, 2003, roč. 18, č. 11, s. 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 a 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, roč.~18, č.~11, s.~1329-1337. ISSN~0956-5663.

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