Stabilization of cholinesterase, detector kit using stabilized cholinesterase, and methods of making and using the same
First Claim
1. A method for providing a treated porous material impregnated with a substantially stabilized cholinesterase comprising the steps of:
- (a) uniformly distributing the cholinesterase through an aqueous medium buffered to a pH greater than about 6 but less than about 9 with a buffering amount of a zwitterionic buffering agent,(b) impregnating a porous material with the aqueous medium containing said cholinesterase, and(c) drying the resulting impregnated porous material to remove substantially all of the physically bound water and thereby provide said treated porous material, having substantially no measurable water vapor pressure in a closed system.
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Accused Products
Abstract
The stability of a cholinesterase (particularly a cholinesterase solution impregnated into a porous material and air-dried) can be improved by: (a) buffering the cholinesterase solution with a zwitterionic buffer, e.g. a buffer having a sulfonic acid group and a protonatable amine group, and, preferably, (b) further drying the impregnated, air-dried porous material under a high vacuum (e.g. 0.01 mm Hg or less) at normal ambient temperatures. The most useful porous materials are sheet-like in nature; that is, they have only two major surfaces. An impregnated, sheet-like material of this invention can be used in a cholinesterase inhibitor detector kit. A typical kit of this type provides a simple means for detecting, inter alia, environmental cholinesterase-inhibiting pollutant, e.g. organophosphorous pesticides and nerve agents.
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Citations
19 Claims
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1. A method for providing a treated porous material impregnated with a substantially stabilized cholinesterase comprising the steps of:
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(a) uniformly distributing the cholinesterase through an aqueous medium buffered to a pH greater than about 6 but less than about 9 with a buffering amount of a zwitterionic buffering agent, (b) impregnating a porous material with the aqueous medium containing said cholinesterase, and (c) drying the resulting impregnated porous material to remove substantially all of the physically bound water and thereby provide said treated porous material, having substantially no measurable water vapor pressure in a closed system. - View Dependent Claims (2, 3, 4, 5, 6, 7, 14)
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8. A method for providing a treated porous material impregnated with a substantially stabilized electric eel cholinesterase comprising the steps of:
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(a) uniformly distributing the cholinesterase through an aqueous medium buffered to a pH greater than about 6 but less than about 9 with a buffering amount of a zwitterionic buffering agent, (b) impregnating a porous material with the aqueous medium containing said cholinesterase, and (c) subjecting the resulting impregnated porous material to a partial vacuum for at least 1 hour in a zone of subatmospheric pressure and generally normal ambient temperature;
the pressure in said zone being less than 1.0 mm of mercury;
thereby obtaining said treated porous material. - View Dependent Claims (9, 10, 11, 12, 13)
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15. A method for providing a treated porous material impregnated with a substantially stabilized electric eel cholinesterase comprising the steps of:
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(a) uniformly distributing the cholinesterase through an aqueous medium buffered to a pH greater than about 6 but less than about 9 with a buffering amount of a zwitterionic buffering agent, (b) impregnating a porous material with the aqueous medium containing said cholinesterase, and (c) subjecting the resulting impregnated porous material to a partial vacuum in a zone of subatmospheric pressure and generally normal ambient temperature;
the pressure in said zone being less than 1.0 mm of mercury;
thereby removing substantially all physically bound water from the resulting dried electric eel cholinesterase, the water vapor pressure of said dried electric eel cholinesterase being substantially unmeasurable in a closed system.
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16. An anti-cholinesterase detector kit, said kit consisting essentially of the following components contained within a moisture resistant package:
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(a) an extremely dry, impregnated porous material, said porous material being impregnated with cholinesterase and a buffering amount of a zwitterionic buffering agent, said impregnated porous material being dried by subjecting the resulting impregnated porous material to a partial vacuum for at least 1 hour in a zone of subatmospheric pressure and generally normal ambient temperature;
the pressure in said zone being less than 1.0 mm of mercury;(b) a dry ester substrate for the cholinesterase, said substrate being separated from the cholinesterase by a barrier for preventing cholinesterase inactivation by sublimation of the substrate during storage. - View Dependent Claims (17, 18, 19)
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Specification