Miles Laboratories, Inc.

Miles Inc.

Greenquist, Alfred C.

Alfred C Greenquist

Bert Walter

Ronald G Sommer

Richard J Tyhach

Patricia A Rupchock

Thomas M Li

A multizone test device for the determination of analyte from a liquid test medium upon contact with the liquid test medium and a labeled reagent comprising a chemical group having a detectable chemical property. The test device preferably comprises multilayers including a reagent layer incorporated with an immobilized reagent and a detection layer incorporated with an immobilized form of an interactive detection reagent for the labeled reagent. The immobilized reagent in the reagent layer and the labeled reagent comprise specific binding partners which will bind to each other dependent upon the amount of analyte present. Labeled reagent which does not become bound to the immobilized reagent in the reagent layer migrates into the detection layer and interacts with the immobilized interactive detection reagent therein which results in the localized generation of a detectable reaction product which preferably is also immobilized in the detection zone. As a result, reverse migration of the labeled reagent, and preferably the detectable reaction product from the detection layer is prevented and the detectable chemical property provided by the label of the labeled reagent is localized in the detection layer for the precise measurement thereof and correlation to the amount of analyte in the test medium.

Multizone analytical element having detectable signal concentrating zone

A multizone test device for the determination of analyte from a liquid test medium upon contact with the liquid test medium and a labeled reagent comprising a chemical group having a detectable physical property. The test device preferably comprises multilayers including a reagent layer incorporated with an immobilized reagent and a detection layer incorporated with an immobilized form of a binding substance for the labeled reagent. The immobilized reagent and the labeled reagent comprise specific binding partners which will bind to each other dependent upon the amount of analyte present. Labeled reagent which does not become bound to the immobilized reagent in the reagent layer migrates into the detection layer and becomes bound to and immobilized by the immobilized binding substance therein. As a result, reverse migration of the labeled reagent into the reagent layer is prevented and the detectable physical property provided by the label of the labeled reagent is localized in the detection layer for the precise measurement thereof and correlation to the amount of analyte in the test medium.

Multizone analytical element having labeled reagent concentration zone

A heterogeneous specific binding assay method for determining the amount of a suspected analyte in an aqueous test medium wherein a reaction mixture is formed by combining the test medium with assay reagents including a labeled reagent, an immobilizable component, and a binding substance which causes the immobilizable component to precipitate. Free and bound species of the labeled reagent are formed as a function of the amount of the analyte in the test medium. One of the free and bound species of the labeled reagent is immobilized by binding of the immobilizable component with the binding substance. The immobilized labeled reagent is seperated from labeled reagent which has not been immobilized, and the amount of label in the labeled reagent in one of the separated fractions is determined and related to the amount of analyte in the test medium. The improvement provided is the use of an immobilizable component comprising a water dispersible, aggregatable reagent comprising a binding partner for the one of the free and bound species to be immobilized and a first binding substance which upon binding of a second binding substance forms a precipitated complex of the aggregatable reagent.

Heterogeneous specific binding assay employing an aggregatable binding

A homogeneous specific binding assay device, a method for its preparation, and a method for its use in determining a ligand, such as antigen, hapten, or antibody, in, or the ligand binding capacity of, a liquid sample. The test device comprises a solid carrier member, such as a fibrous web matrix, e.g., paper, or a polymeric film or gel, incorporated with reagents for a homogeneous specific binding assay system which produces a detectable response, usually an electromagnetic radiation signal, that is a function of the presence or amount of the ligand in or the ligand binding capacity of the sample. Useful homogeneous specific binding assay systems include those involving enzyme substrate labels, enzyme prosthetic group labels, and enzyme labels. The detectable response preferably is a luminescent, fluorescent, spectrophotometric, or colorimetric response, which is measurable by visual observation or instrument means.

Multilayer homogeneous specific binding assay device

A device for the quantitative determination of heparin in mammalian blood plasma, a method for the preparation of such device and a method for determining the plasma's heparin concentration using the device. The device, which is prepared using a freeze-drying technique, comprises a first upper layer of a carrier matrix material containing a buffer, AT-III and thrombin and a second, lower layer of a carrier matrix containing a thrombin sensitive fluorogenic or chromogenic substrate being capable of interacting with thrombin in such a manner that a time-related chemical change detectable by fluorometric or spectrophotometric means takes place when thrombin and the substrate are contacted in a suitable liquid environment.

Device and method for the quantitative determination of heparin in

Disclosed is a method and device for the quantitative determination of AT-III in mammalian blood plasma. The method involves contacting a plasma sample containing excess heparin with a 3 layered reagent strip comprising:

Reagent strip test for antithrombin-III

A homogeneous specific binding assay element, a method for its preparation, and a method for its use in determining a ligand in or the ligand binding capacity of a liquid sample are disclosed. The test element comprises a solid carrier incorporated with reagents for a homogeneous specific binding assay system which produces a detectable response, usually an electromagnetic radiation signal, that is a function of the presence or amount of the ligand in or the ligand binding capacity of the sample. There is further disclosed a method for preparing a homogeneous specific binding assay element for determining a ligand in or the ligand binding capacity of a liquid sample by incorporating a carrier with a composition which includes a label conjugate, comprising a label component coupled to a ligand moiety or a specific binding analog thereof, and a reagent reactive with the label conjugate, which method comprises (a) incorporating the carrier with the reagent reactive with the label conjugate in a first liquid; (b) subjecting the carrier of (a) to conditions effective to reversibly suspend or reduce the activity of the reagent therein; (c) incorporating the carrier of (b) with the label conjugate; (d) subjecting the carrier of (c) to a temperature effective to freeze the reagent reactive with the label conjugate and the label conjugate; and (e) lyophilizing the reagent and label conjugate in the carrier of (d).

Homogeneous specific binding assay element and lyophilization production

A homogeneous specific binding assay element, a method for its preparation, and a method for its use in determining a ligand in or the ligand binding capacity of a liquid sample. The test element comprises a solid carrier incorporated with reagents for a homogeneous specific binding assay system which produces a detectable response, usually an electromagnetic radiation signal, that is a function of the presence or amount of the ligand in or the ligand binding capacity of the sample. Also a method for preparing a homogeneous specific binding assay element for determining a ligand in or the ligand binding capacity of a liquid sample by incorporating a carrier with a composition which includes a label conjugate, comprising a label component coupled to a ligand moiety or a specific binding analog thereof, and a reagent reactive with the label conjugate, which method comprises (a) incorporating the carrier with the reagent reactive with the label conjugate in a first liquid and drying the carrier; and then (b) incorporating the carrier of (a) with the label conjugate in a liquid effective to prevent reaction with the reagent reactive with the label conjugate prior to contact of the element with the sample and drying the carrier.

Preparing homogeneous specific binding assay element to avoid premature

A homogeneous specific binding assay device, a method for its preparation, and a method for its use in determining a ligand, such as antigen, hapten, or antibody, in a liquid sample. The test device comprises a solid carrier member, such as a fibrous web matrix, e.g., paper, or a polymeric film or gel, incorporated with reagents for a homogeneous specific binding assay system which produces a detectable response, usually an electromagnetic radiation signal, that is a function of the presence or amount of the ligand in the sample. For example, disclosed is a test device for determining a ligand in a liquid sample, comprising (a) a reagent composition including a complex of (i) a labeled conjugate comprising a label component coupled to said ligand or a specific binding analog thereof, and (ii) a specific binding partner for said ligand, said label providing a detectable response, or interacting with a detectant system to provide a detectable response, which is different when the labeled conjugate is bound by said binding partner compared to when it is not so bound, whereby the detectable response is a function of the presence of the ligand in the sample, and (b) a carrier incorporated with said complex. Useful homogeneous specific binding assay systems include those involving enzyme substrate labels, enzyme prosthetic group labels, and enzyme labels. The detectable response preferably is a luminescent, fluorescent, spectrophotometric, or colorimetric response, which is measurable by visual observation or instrument means.

Homogeneous specific binding assay device and preformed complex method

A multilayer analytical element for detecting a ligand in or the ligand binding capacity of a liquid sample of the type having a reagent layer(s) incorporating reagents which are responsive to the ligand in or the ligand binding capacity of the sample to give a detectable response, a radiation reflecting layer, and a support layer, the improvement wherein the reagent layer(s) is a radiation diffusing layer and the radiation reflecting layer is a radiation nondiffusing reflecting layer which is (a) interposed between the reagent layer(s) and the support layer; (b) impermeable to the ligand, reagents of the reagent layer(s), and products of their interreaction; and (c) inert to the ligand, reagents of the reagent layer(s), and products of their interreaction.

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Elkhart, IN, United States of America

Alfred C Greenquist