Development of a bifunctional sensor using haptenized acetylcholinesterase and application for the detection of cocaine and organophosphates

Carsten Teller, J. Halamek, Jiri Žeravík, Walter F.M. Stöcklein, Frieder W. Scheller

Research output: Contribution to journalArticleAcademic

12 Citations (Scopus)

Abstract

We developed a dual piezoelectric/amperometric sensor for the detection of two unrelated analytes in one experiment that uses propidium to anchor acetylcholinesterases (AChE) at the surface. This mass-sensitive sensor does not only allow the examination of the interaction between AChE and the modified surface but also the detection of in situ inhibition of the surface-bound AChE. Here we describe the application of the propidium-based sensor in combination with a modified AChE. For this reason the cocaine derivative benzoylecgonine (BZE) was coupled via a 10 Å long hydrophilic linker – 1,8-diamino-3,4-dioxaoctane – to carboxylic groups of the AChE after EDC/NHS activation. Thus the modified AChE (BZE–AChE) possesses an additional recognition element besides the inhibitor binding site. After the deposition of BZE–AChE on the sensor surface the binding of an anti-BZE-antibody to the BZE–AChE can be monitored. This makes it possible to determine two analytes – cocaine and organophosphate – in one experiment by measuring antibody binding and decrease in enzymatic activity, respectively. Furthermore it was also shown that other cocaine-binding enzymes, e.g., butyrylcholinesterase, can bind to the modified BZE–AChE. The competitive immunoassay allowed the detection of cocaine with a dynamic range from 10−9 to 10−7 M. The organophosphate chlorpyrifos-oxon could be detected in concentrations from 10−6 down to 10−8 M after 20 min of injection time (equals to 500 L sample volume.
Original languageUndefined
Pages (from-to)111-117
JournalBiosensors and bioelectronics
Volume24
Issue number1
DOIs
Publication statusPublished - 2008

Keywords

  • Detection
  • Modified enzyme
  • Cocaine
  • Organophosphate
  • Acetylcholinesterase
  • IR-79100
  • Propidium

Cite this

Teller, Carsten ; Halamek, J. ; Žeravík, Jiri ; Stöcklein, Walter F.M. ; Scheller, Frieder W. / Development of a bifunctional sensor using haptenized acetylcholinesterase and application for the detection of cocaine and organophosphates. In: Biosensors and bioelectronics. 2008 ; Vol. 24, No. 1. pp. 111-117.
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abstract = "We developed a dual piezoelectric/amperometric sensor for the detection of two unrelated analytes in one experiment that uses propidium to anchor acetylcholinesterases (AChE) at the surface. This mass-sensitive sensor does not only allow the examination of the interaction between AChE and the modified surface but also the detection of in situ inhibition of the surface-bound AChE. Here we describe the application of the propidium-based sensor in combination with a modified AChE. For this reason the cocaine derivative benzoylecgonine (BZE) was coupled via a 10 {\AA} long hydrophilic linker – 1,8-diamino-3,4-dioxaoctane – to carboxylic groups of the AChE after EDC/NHS activation. Thus the modified AChE (BZE–AChE) possesses an additional recognition element besides the inhibitor binding site. After the deposition of BZE–AChE on the sensor surface the binding of an anti-BZE-antibody to the BZE–AChE can be monitored. This makes it possible to determine two analytes – cocaine and organophosphate – in one experiment by measuring antibody binding and decrease in enzymatic activity, respectively. Furthermore it was also shown that other cocaine-binding enzymes, e.g., butyrylcholinesterase, can bind to the modified BZE–AChE. The competitive immunoassay allowed the detection of cocaine with a dynamic range from 10−9 to 10−7 M. The organophosphate chlorpyrifos-oxon could be detected in concentrations from 10−6 down to 10−8 M after 20 min of injection time (equals to 500 L sample volume.",
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Development of a bifunctional sensor using haptenized acetylcholinesterase and application for the detection of cocaine and organophosphates. / Teller, Carsten; Halamek, J.; Žeravík, Jiri; Stöcklein, Walter F.M.; Scheller, Frieder W.

In: Biosensors and bioelectronics, Vol. 24, No. 1, 2008, p. 111-117.

Research output: Contribution to journalArticleAcademic

TY - JOUR

T1 - Development of a bifunctional sensor using haptenized acetylcholinesterase and application for the detection of cocaine and organophosphates

AU - Teller, Carsten

AU - Halamek, J.

AU - Žeravík, Jiri

AU - Stöcklein, Walter F.M.

AU - Scheller, Frieder W.

PY - 2008

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N2 - We developed a dual piezoelectric/amperometric sensor for the detection of two unrelated analytes in one experiment that uses propidium to anchor acetylcholinesterases (AChE) at the surface. This mass-sensitive sensor does not only allow the examination of the interaction between AChE and the modified surface but also the detection of in situ inhibition of the surface-bound AChE. Here we describe the application of the propidium-based sensor in combination with a modified AChE. For this reason the cocaine derivative benzoylecgonine (BZE) was coupled via a 10 Å long hydrophilic linker – 1,8-diamino-3,4-dioxaoctane – to carboxylic groups of the AChE after EDC/NHS activation. Thus the modified AChE (BZE–AChE) possesses an additional recognition element besides the inhibitor binding site. After the deposition of BZE–AChE on the sensor surface the binding of an anti-BZE-antibody to the BZE–AChE can be monitored. This makes it possible to determine two analytes – cocaine and organophosphate – in one experiment by measuring antibody binding and decrease in enzymatic activity, respectively. Furthermore it was also shown that other cocaine-binding enzymes, e.g., butyrylcholinesterase, can bind to the modified BZE–AChE. The competitive immunoassay allowed the detection of cocaine with a dynamic range from 10−9 to 10−7 M. The organophosphate chlorpyrifos-oxon could be detected in concentrations from 10−6 down to 10−8 M after 20 min of injection time (equals to 500 L sample volume.

AB - We developed a dual piezoelectric/amperometric sensor for the detection of two unrelated analytes in one experiment that uses propidium to anchor acetylcholinesterases (AChE) at the surface. This mass-sensitive sensor does not only allow the examination of the interaction between AChE and the modified surface but also the detection of in situ inhibition of the surface-bound AChE. Here we describe the application of the propidium-based sensor in combination with a modified AChE. For this reason the cocaine derivative benzoylecgonine (BZE) was coupled via a 10 Å long hydrophilic linker – 1,8-diamino-3,4-dioxaoctane – to carboxylic groups of the AChE after EDC/NHS activation. Thus the modified AChE (BZE–AChE) possesses an additional recognition element besides the inhibitor binding site. After the deposition of BZE–AChE on the sensor surface the binding of an anti-BZE-antibody to the BZE–AChE can be monitored. This makes it possible to determine two analytes – cocaine and organophosphate – in one experiment by measuring antibody binding and decrease in enzymatic activity, respectively. Furthermore it was also shown that other cocaine-binding enzymes, e.g., butyrylcholinesterase, can bind to the modified BZE–AChE. The competitive immunoassay allowed the detection of cocaine with a dynamic range from 10−9 to 10−7 M. The organophosphate chlorpyrifos-oxon could be detected in concentrations from 10−6 down to 10−8 M after 20 min of injection time (equals to 500 L sample volume.

KW - Detection

KW - Modified enzyme

KW - Cocaine

KW - Organophosphate

KW - Acetylcholinesterase

KW - IR-79100

KW - Propidium

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DO - 10.1016/j.bios.2008.03.027

M3 - Article

VL - 24

SP - 111

EP - 117

JO - Biosensors and bioelectronics

JF - Biosensors and bioelectronics

SN - 0956-5663

IS - 1

ER -