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  • Complex Coacervate Core Micelles for the Dispersion and Stabilization of Organophosphate Hydrolase in Organic Solvents.

Complex Coacervate Core Micelles for the Dispersion and Stabilization of Organophosphate Hydrolase in Organic Solvents.

Langmuir : the ACS journal of surfaces and colloids (2016-12-21)
Carolyn E Mills, Allie Obermeyer, Xuehui Dong, Jeremy Walker, Bradley D Olsen
ABSTRACT

Organophosphate (OP) nerve agents are a class of chemical warfare agents (CWAs) that exist as bulk stocks in current and past war zones. Thus, a technology that can perform on-site decontamination in a safe and timely fashion is desirable. Here, complex coacervate core micelles (C3Ms) were used to encapsulate organophosphate hydrolase (OPH) and chemostabilize it to maintain activity after exposure to organophosphate simulants ethanol and dimethyl methylphosphonate (DMMP). C3Ms were formed by two polymers-poly(acrylic acid) (PAA) and poly(oligo(ethylene glycol) methacrylate)-b-poly(4-vinyl N-methylpyridyl iodide), (POEGMA-b-qP4VP). Complexes of the coacervate micelles with the enzyme OPH were investigated by small angle neutron scattering (SANS), dynamic light scattering (DLS), and transmission electron microscopy (TEM), demonstrating the formation of micellar structures in solution. The activity of OPH against methyl paraoxon in these C3Ms under aqueous conditions was assayed after heat treatment for 3 days at 37 °C. The OPH in C3Ms retained 88 ± 7% of its initial activity, as compared to the 48 ± 3% activity retained by OPH alone, indicating that the C3Ms were able to stabilize the enzyme to heat treatment. C3Ms transferred into the two organic solvents formed larger structures than inverse micelles formed by the block copolymer alone. The addition of OPH to the C3Ms in organic solvents did not significantly change their structure. The activity of OPH (again, against methyl paraoxon) after 24 h of incubation at 4 °C was measured and compared to that of OPH in C3Ms. While OPH alone retained less than 5% of its activity after this incubation in both solvents, OPH in C3Ms retained 35 ± 3% of its activity in DMMP and 26 ± 1% of its activity in ethanol.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
α-Chymotrypsinogen A from bovine pancreas, essentially salt-free, lyophilized powder
Sigma-Aldrich
Iodomethane, contains copper as stabilizer, ReagentPlus®, 99%
Sigma-Aldrich
4-Cyano-4-(phenylcarbonothioylthio)pentanoic acid
Sigma-Aldrich
4-Vinylpyridine, contains 100 ppm hydroquinone as inhibitor, 95%
Sigma-Aldrich
Poly(acrylic acid sodium salt), average Mw ~5,100 by GPC, powder