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 (276 Německo), Jan HALÁMEK (203 Česká republika), Petr SKLÁDAL (203 Česká republika, garant), Franz KERNCHENC (276 Německo) a Frieder SCHELLER (276 Německo)
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
Kód RIV
RIV/00216224:14310/03:00008862
Organizační jednotka
Přírodovědecká fakulta
Klíčová slova anglicky
cholinesterase; inhibitor; piezoelectric; biosensor; real-time
Štítky
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 |
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