Campus Units

Mechanical Engineering

Document Type

Article

Publication Version

Published Version

Publication Date

2017

Journal or Book Title

Analyst

Volume

142

First Page

3261

Last Page

3271

DOI

10.1039/C6AN02575G

Abstract

The rapid detection of organophosphates (OPs), a class of strong neurotoxins, is critically important for monitoring acute insecticide exposure and potential chemical warfare agent use. Herein, we improve the enzymatic activity of a phosphotriesterase trimer (PTE3), an enzyme that selectively recognizes OPs directly, by conjugation with distinctly sized (i.e., 5, 10, and 20 nm diameter) gold nanoparticles (AuNPs). The number of enzymes immobilized on the AuNP was controlled by conjugating increasing molar ratios of PTE3 onto the AuNP surface via metal affinity coordination. This occurs between the PTE3-His6 termini and the AuNP-displayed Ni2+-nitrilotriacetic acid end groups and was confirmed with gel electrophoresis. The enzymatic efficiency of the resultant PTE3–AuNP bioconjugates was analyzed via enzyme progress curves acquired from two distinct assay formats that compared free unbound PTE3 with the following PTE3–AuNP bioconjugates: (1) fixed concentration of AuNPs while increasing the bioconjugate molar ratio of PTE3 displayed around the AuNP and (2) fixed concentration of PTE3 while increasing the bioconjugate molar ratio of PTE3–AuNP by decreasing the AuNP concentration. Both assay formats monitored the absorbance of p-nitrophenol that was produced as PTE3 hydrolyzed the substrate paraoxon, a commercial insecticide and OP nerve agent simulant. Results demonstrate a general equivalent trend between the two formats. For all experiments, a maximum enzymatic velocity (Vmax) increased by 17-fold over free enzyme for the lowest PTE3–AuNP ratio and the largest AuNP (i.e., ratio of 1[thin space (1/6-em)]:[thin space (1/6-em)]1, 20 nm dia. AuNP). This work provides a route to improve enzymatic OP detection strategies with enzyme–NP bioconjugates.

Comments

This article is published as Hondred, John A., Joyce C. Breger, Nate T. Garland, Eunkeu Oh, Kimihiro Susumu, Scott A. Walper, Igor L. Medintz, and Jonathan C. Claussen. "Enhanced enzymatic activity from phosphotriesterase trimer gold nanoparticle bioconjugates for pesticide detection." Analyst 142, no. 17 (2017): 3261-3271. DOI: 10.1039/C6AN02575G . Posted with permission.

Rights

Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.

Language

en

File Format

application/pdf

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