Optical oxidative enzyme-based sensors
First Claim
1. An oxidative enzyme sensor having an optical sensing membrane comprising, in order:
- (a) an enzymatic layer containing an oxidative enzyme in a hydrateable and oxygen permeable matrix;
(b) an oxygen sensing layer containing luminescent dye in a light-transmissive, oxygen permeable matrix wherein said enzymatic layer is disposed on said oxygen sensing layer, and(c) a rapidly hydrating gas-permeable cover layer disposed over the enzymatic layer, wherein the cover layer is effective to delay oxygen released by red blood cells from reaching the oxygen sensing layer for a period of time.
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Abstract
The invention discloses enzyme-based optical sensors for detecting blood components which are substrates for oxidative enzymes, the sensors advantageously employing a multiple-layer structure featuring a thin, rapidly responding, optical, oxygen sensing layer. The sensors comprise, in order, a) an enzymatic layer containing an oxidative enzyme or enzyme cascade in a water and oxygen-permeable matrix; b) an oxygen sensing layer containing luminescent dye in a light-transmissive, oxygen-permeable matrix; and which is preferably deposited onto c) a light-transmissive substrate. Embodiments of the invention may further include a rapidly hydrating gas-permeable cover, or spacer, layer deposited over the enzymatic layer. A particulate filler material may be included in an effective amount in the oxygen sensing layer to reduce sample light scattering effects. The sensors of the disclosure may be used to detect creatinine and other enzyme-oxidizable analytes such as glucose, lactate or cholesterol, and may be made easily using standard coating techniques known in the art. These sensors are suitable for multiple analyses, but the materials and methods used allow the sensors to be disposable as well, allowing their wide use in medical and analytical applications.
535 Citations
22 Claims
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1. An oxidative enzyme sensor having an optical sensing membrane comprising, in order:
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(a) an enzymatic layer containing an oxidative enzyme in a hydrateable and oxygen permeable matrix; (b) an oxygen sensing layer containing luminescent dye in a light-transmissive, oxygen permeable matrix wherein said enzymatic layer is disposed on said oxygen sensing layer, and (c) a rapidly hydrating gas-permeable cover layer disposed over the enzymatic layer, wherein the cover layer is effective to delay oxygen released by red blood cells from reaching the oxygen sensing layer for a period of time. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 17, 18, 19)
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10. A method of making an oxidative enzyme sensor having an optical sensing membrane comprising in order:
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(a) preparing an oxygen sensing layer containing a luminescent dye in a light-transmissive, oxygen permeable matrix; (b) depositing an enzymatic layer on said oxygen sensing layer, said enzymatic layer containing an oxidative enzyme or oxidative enzyme cascade in an oxygen-permeable matrix; and (c) depositing a rapidly hydrating gas-permeable cover layer on the enzymatic layer, wherein the cover layer is effective to delay oxygen released by red blood cells from reaching the oxygen sensing layer for a period of time. - View Dependent Claims (11, 12, 13, 14, 15, 16, 20, 21, 22)
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Specification