Abstract: Disclosed is a test cell and a method for detection of a preselected ligand in a liquid sample such as a body fluid. The test cell includes an elongate outer casing which houses an interior permeable material capable of transporting an aqueous solution and defining a sample inlet, a test volume, and a reservoir volume. The reservoir volume is disposed in a section of the test cell spaced apart from the inlet and is filled with sorbent material. The reservoir acts to receive liquid transported along a flow path defined by the permeable material and extending from the inlet and through the test volume. In the test volume is a test site which includes a first protein having a binding site specific to a first epitope of the ligand immobilized in fluid communication with the flow path. The test site can be observed through a window of the casing.

Abstract: A prenatal screening method for Down's syndrome involves assaying for hyperglycosylated gonadotropin in biological test samples such as urine, plasma or serum obtained from pregnant women. Hyperglycosylated gonadotropin comprises a variant population of chorionic gonadotropin, chorionic gonadotropin-free &bgr;-subunit, &bgr;-core fragment, and/or free &agr;-subunit exhibiting differences in the carbohydrate content from what is observed in samples obtained from pregnant women carrying normal fetuses. Qualitative or quantitative observation of differences in the carbohydrate content of the hyperglycosylated gonadotropin population from corresponding control samples containing a normal gonadotropin population, or direct observation of the variant species seen in Down's syndrome, indicates that the woman's fetus has Down's syndrome. Typical screens involve carbohydrate analyses, immunoassays, or combinations of these methods.

Abstract: The present invention provides a novel chemiluminescent reagent producing chemiluminescence in the presence of hydrogen peroxide, extent of which depends on peroxidase concentration, chemiluminescent analysis method using the same, in particular useful for detection and quantitative analysis of various types of materials by measuring peroxidase enzyme activity or enzyme immunoassay with peroxidase enzyme as the marker. More particularly, the present invention provides a chemiluminescent reagent containing, as the major ingredients, a charge-transferring complex of N,N′-disubstituted-9,9′-bisacridinium salt and N,N-disubstituted carboxylic amide compound; chemiluminescent reagent containing further containing a specific aminoalcohol compound, in addition to the above; and method for measuring peroxidase activity at a high sensitivity in the presence of a peroxide, using the above chemiluminescent reagent.

Abstract: A solution of a non-reconstituted hormone and a preservative having a useful shelf life of at least six months, and, in some circumstances, at least a year. A series of solution of varying levels of the non-reconstituted hormone may be in a test kit and used in a process to determine the level of the hormone in a patient sample. In an aspect of the invention, the hormone is parathyroid hormone. The preservatives may be polyvinyl alcohol, a dissolved EDTA salt, or dissolved sodium molybdate. The hormone may be in an aqueous or a buffered matrix containing very little protein and substantial amounts of TX100 to reduce non-specific binding. The solution is incubated with a solid phase coated with an anti-hormone antibody and a labeled hormone antibody. The solid phase is read using a suitable device for measuring the labeled hormone antibody. An example of a labeled hormone antibody is an acridinium labeled hormone antibody that is read using a light reading device.

Abstract: The present invention relates to compositions comprising novel reduced cortisol conjugates, methods for their preparation and use in immunoassays for cortisol. In another aspect, it relates to conjugates of reduced cortisol as immunogens or haptens for eliciting anti-cortisol or anti-reduced cortisol antibodies.

Abstract: A method and compositions for the detection and/or quantification of an analyte through the use of a plurality of labeled probes, with two or more said probes targeted to different regions of said analyte. In specific embodiments, the labels are separately distinguishable, and/or are present at different specific activities on the labels.

Abstract: Receptors are disclosed that are antibodies that exhibit a binding affinity for an immune complex of a monoepitopic antigen and an antibody for such antigen that is substantially greater than the binding affinity for the monoepitopic antigen or the antibody for the monoepitopic antigen apart from the immune complex. Normally, the monoepitopic antigen has a molecular weight less than 1500 and is an organic compound. The antibodies of the present invention find use in a method for determining a monoepitopic antigen in a sample suspected of containing such antigen. The method comprises forming an immune sandwich complex comprising the monoepitopic antigen or an analog thereof, a first monoclonal antibody that binds to the monoepitopic antigen, and a second monoclonal antibody that is an antibody of the present invention and detecting the immune sandwich complex.

Abstract: The present invention provides an improved method for using the measurement of total urinary 17 KS-S, referred to herein as the urinary Anabolic/Catabolic Index (ACI), as part of a biomarker for biological age.

Abstract: The present invention relates to a monoclonal antibody binding an antigen on the megakaryocytic cell line UT-7. The invention further relates to hybridoma cells producing such an antibody.

Abstract: A method and apparatus for measuring binding between a plurality of molecules of a biological receptor protein and a plurality of molecules of a type which binds to said biological receptor is presented. Apparatus utilizes a sensor possessing a waveguide to which have been attached in close proximity to its surface, features resembling molecules having binding affinity for said biological receptor. Light is injected into said waveguide so as to produce an evanescent field at its surface. Molecules of receptor protein are tagged with a tag belonging to that class of chemicals which alters a characteristic of light, when said light passes through said chemical tag. Apparatus utilizes a rapid means of monitoring the change in optical signal coming from the waveguide as binding proceeds between tagged receptor protein and the binding molecular feature of the waveguide. This allows direct measurement of binding and dissociation rates between the receptor and the binding feature of the waveguide.

Abstract: Calibrating an immunoassay by generating two reaction rate measuring curves, from samples having higher and lower relative levels of antigen, extrapolating a combination of the curves to cover sample concentrations known to contain an excess of antigen relative to an amount of capture reagent and combining the low end linear potion of the higher reaction rate measuring curve with the higher end portion of the extrapolated reaction rate measuring curve, thereby eliminating measuring inaccuracies otherwise arising from the hook effect. For antigen concentrations higher than the assay range, a high antigen signal utilizing the two rates avoids reporting false results.

Abstract: A method and apparatus for measuring binding between a plurality of molecules of a first type and a plurality of molecules of a second type is presented. Apparatus utilizes a sensor possessing a waveguide to which have been attached in close proximity to its surface, features resembling molecules of said first type. Light is injected into said waveguide so as to produce an evanescent field at its surface. Molecules of said second type are tagged with a tag belonging to that class of chemicals which alters a characteristic of light, when said light passes through said chemical tag. Apparatus utilizes a rapid means of monitoring the change in optical signal coming from said waveguide as binding proceeds between tagged molecules of type 2 and the feature resembling molecules of type 1 on said waveguide. This allows direct measurement of binding and dissociation rates between the two types of molecules.

Abstract: An analytical test device useful for example in pregnancy testing, comprises a hollow casing (500) constructed of moisture-impervious solid material, such as plastics materials, containing a dry porous carrier (510) which communicates indirectly with the exterior of the casing via a bibulous sample receiving member (506) which protrudes from the casing such that a liquid test sample can be applied to the receiving member and permeate therefrom to the porous carrier, the carrier containing in a first zone a labelled specific binding reagent is freely mobile within the porous carrier when in the moist state, and in a second zone spatially distinct from the first zone unlabelled specific binding reagent for the same analyte which unlabelled reagent is permanently immobilized on the carrier material and is therefore not mobile in the moist state, the two zones being arranged such that liquid sample applied to the porous carrier can permeate via the first zone into the second zone, and the device incorporating mea

Abstract: The invention deals with the use of a reactive compound in immunological analyses. The compound is ouabain or its analog to which another compound, labeled by radioiodine or by a fluorogen, is coupled. The invention covers also the method to measure ouabain or its analog in plasma to diagnose cardiovascular and endocrine diseases and other harmful conditions.

Abstract: An analytical test device useful for example in pregnancy testing, comprises a hollow casing (500) constructed of moisture-impervious solid material, such as plastics materials, containing a dry porous carrier (510) which communicates indirectly with the exterior of the casing via a bibulous sample receiving member (506) which protrudes from the casing such that a liquid test sample can be applied to the receiving member and permeate therefrom to the porous carrier, the carrier containing in a first zone a labelled specific binding reagent is freely mobile within the porous carrier when in the moist state, and in a second zone spatially distinct from the first zone unlabelled specific binding reagent for the same analyte which unlabelled reagent is permanently immobilised on the carrier material and is therefore not mobile in the moist state, the two zones being arranged such that liquid sample applied to the porous carrier can permeate via the first zone into the second zone, and the device incorporating mea

Abstract: A method and compositions for the detection and/or quantification of an analyte through the use of a plurality of labeled probes, with two or more said probes targeted to different regions of said analyte. In specific embodiments, the labels are separately distinguishable, and/or are present at different specific activities on the labels.

Abstract: A new class of chemiluminescent acridinium or benzacridinium compounds is disclosed by virtue of forming an intramolecular energy transfer conjugate (ETC) between the acridinium or benzacridinium compound and a luminophore. A method of extending the emission wavelengths of acridinium or benzacridinium esters in order to further reduce or eliminate the emission spectral overlap between the parent polysubstituted aryl Acridinium Esters (DMAE) and Benzacridinium Esters (LEAE) is disclosed. The ETC's retain the unique desired properties of acridinium or benzacridinium compounds including complete light emission in very short period of time, monophasic emission spectrum, simplicity of triggering mechanism, ability of labeling the biological molecules of interest to form a tracer, and good stability. Additionally, the range of the emission spectrum of an acridinium or benzacridinium compound can now be shifted at will and at longer leap through the choice of a luminophore as the integral part of an ETC molecule.

Abstract: The present invention relates to compounds that are bis-biotins. These compounds comprise two biotinyl radicals connected by a chain of atoms, usually at least 16 atoms in length. The bis-biotin is conjugated to a member of a specific binding pair ("sbp member") wherein the chain is not part of the sbp member. Also disclosed are compositions comprising a complex of avidin and a bis-biotin as described above. The compounds and compositions of the invention find use in an assay for an analyte wherein there is employed a reagent system comprising an avidin reagent and a biotin reagent. The improvement of the present invention comprises using as the biotin reagent a bis-biotin as described above. Also disclosed are kits comprising the present bis-biotins and methods of preparing a bis-biotinylated conjugate of a member of a specific binding pair ("sbp member") for use in a specific binding assay.

Abstract: The present invention is directed to fluorescent dyes and its valence tautomers of the formula wherein Q represents a conjugated moiety that increases the fluorescent quantum yield of the compound; R.sub.1 is a functionalized group of the formula--(CH.sub.2).sub.j Y, wherein Y is selected from the group consisting of SO.sub.3 H, COOH, NH.sub.2, CHO, NCS, epoxy, phthalimido, and COOZ, wherein Z represents a leaving group; R.sub.2 is a functionalized group of the formula --(CH.sub.2).sub.k Y', wherein Y' is selected from the group consisting of SO.sub.3 H, COOH, NH.sub.2, CHO, NCS, epoxy, phthalimido, and COOZ, wherein Z represents a leaving group; M.sup.+ is a counterion selected from the group consisting of ammonium, alkali metal cations, and alkaline earth metal cations; n=1 to 4; m=1 to 4; j=2 to 10; and k=2 to 10.

Abstract: There are provided methods for assaying biological specimens for one or more of leptin, prorenin or renin in order to provide predictive information about the likelihood of a woman carrying a Downs Syndrome affected fetus.

Abstract: A method of diagnosing cardiac transplant rejection within a patient comprising, obtaining a sample of a biological fluid from the patient, and determining the level of a brain natriuretic peptide (BNP) or a fragment thereof, within the sample of body fluid. The step of determining the concentration of BNP involves an assay comprising at least one antibody exhibiting affinity for the BNP or a fragment thereof, and the biological fluid comprises plasma, urine or cerebrospinal fluid. Furthermore, the antibody used within the method may comprises a polyclonal antibody, a monoclonal antibody, or a combination thereof. Preferably, the method involves obtaining at least two of the samples of body fluid from the patient over a period of time and comparing the BNP levels, with an increase in BNP being indicative of an upcoming rejection episode.

Abstract: Fluorescence immunoassays methods are provided which use fluorescent dyes which are free of aggregation and serum binding. Such immunoassay methods are thus, particularly useful for the assay of biological fluids, such as serum, plasma, whole blood and urine.

Abstract: Dimeric inhibin concentration can be used in the determination of the risk of a women carrying an unborn child affected by chromosomal abnormalities, e.g. Downs Syndrome. A method for antenatal screening for chromosomal abnormalities is carried out by measuring body fluid taken from a pregnant woman for the level of at least one marker or precursor or metabolite of the marker together with the gestational age of the woman are compared with reference values taken from women carrying fetuses with chromosomal abnormalities and/or women carrying normal fetuses, using dimeric inhibin as a marker. A second marker is the free beta subunit of human chorion gonadotropin. An apparatus for carrying out the method described comprises a computer with a means for comparing the values described.

Abstract: A method for antenatal screening for chromosomal abnormalities in which maternal blood from a pregnant woman is measured for levels of free beta hCG and at least a second serum marker and/or precursors and metabolites of these markers and the measured levels of these markers together with the gestational age of the pregnant woman are compared to reference values at various gestational ages of the levels for free beta hCG and the second serum marker in (a) pregnant women carrying foetuses having the abnormalitie(s) subject to the screen and (b) pregnant women carrying normal foetuses, the comparison being indicative of the risk of the pregnant woman carrying a foetus with an abnormality subject to the screen characterised in that the second serum marker is pregnancy associated plasma protein A (PAPPA) and the screen is carried out by the end of the thirteenth (13th) completed week of pregnancy.

Abstract: A method for evaluating the solubility capacity of free unbound estrogens in body fluids of animals is described.

Abstract: The present invention relates to methods for measuring endogenous cytokines in blood. The method accurately measures the cytokines in the blood in the presence of substances that bind the cytokines thereby causing the measurement of the cytokines by conventional methods to give inaccurate results. The present invention also includes the non-invasive measurement of cytokines in biological fluids such as saliva and nasal secretions. Finally, the present invention allows one to monitor the level of cytokines in the blood during treatment of a human or animal with cytokines.

Abstract: A method of preventing false detection of signal due to splashing of reagent liquid used to produce such signal, when dispensing at least one such liquid from a metering tip into a second liquid, comprising the steps of: a) positioning the metering tip a predetermined distance above the upper level of the second liquid prior to dispensing the one liquid; andb) while maintaining the distance throughout the dispensing of the one liquid, dispensing the one liquid;wherein the predetermined distance is between about 1.0 mm and about 2.0 mm so that splashing during dispensing is reduced.

Abstract: A method and apparatus for evaluating estrogen dependent physiological conditions includes a substrate having an anthocyanin pigment applied thereto. The pigment yields a color response indicative of how the body fluid responds to changes in its capacity to absorb free estrogens. The pigment is contacted by the body fluid sample, and the response thereto is connected through color response or optical density measurements to predetermined physiological conditions.

Abstract: The present invention is directed to the diagnosis and assessment of wound status by quantitating levels of cortisol present in wound fluid. The present invention is also directed to a kit and test strip for assessing wound status.

Abstract: A device for performing an enzyme-labelled binding assay comprises an absorbent material and a developing solution containing a substrate for said enzyme, wherein the absorbent material is provided with a plurality of reagent zones including an indicator reagent zone and is capable of transporting the developing solution by capillary action sequentially through the reagent zones, and wherein the indicator reagent zone includes a reagent capable, directly or indirectly, of immobilizing an enzyme-labelled reagent in an amount dependent on the assay result, characterized in that the absorbent material includes a reagent that prevents a signal formation except where enzyme-labelled reagent is immobilized at the indicator reagent zone. The absorbent material is suitably in the form of an elongate strip provided with transverse reagent zones. The device is useful for performing immunoassays, including immunometric assays and dual analyte assays.

Abstract: The present invention discloses novel immunogens, antibodies prepared from such immunogens, and labeled reagents useful in immunoassays for the detection and quantification of testosterone in a test sample. Also disclosed are immunoassays using these reagents and methods for synthesizing these reagents. The immunoassays are preferably microparticle enzyme immunoassays (MEIAs) and fluorescence polarization immunoassays (FPIAs). Further disclosed are novel starting materials for making the above novel immunogens and labeled reagents. Methods for making the novel immunogens and labeled reagents from the novel starting materials are also disclosed.

Abstract: A method for making a preconjugate which includes immunogenic species of a polymorphic analyte. The method is carried out by reacting an activated binding moiety, and a polymorphic analyte at room temperature for between about 10 hours and about 60 hours. The attaching reaction results in an excess of the preconjugate which includes the immunogenic species of the polymorphic analyte. The preconjugate can be used to make an immunoreactive conjugate useful as a developer antigen in a competitive inhibition immunoassay for the polymorphic analyte.

Abstract: Fluorescence immunoassays methods are provided which use fluorescent dyes which are free of aggregation and serum binding. Such immunoassay methods are thus, particularly useful for the assay of biological fluids, such as serum, plasma, whole blood and urine.

Abstract: The present invention provides a more accurate risk assessment of antenatal screening results because the maternal age of the patient is considered in evaluating the results from various common screening markers, and especially the results from UE determinations.

Abstract: The present invention provides devices and methods for the detection of creatinine in biological fluids using lateral flow methodologies. The invention is particularly useful in providing one step creatinine assays for correcting urinary steroid hormone assays.

Abstract: The present invention relates to methods for diagnosing acute myocardial infarction through the measurement of the level of marinobufagin-like immunoreactivity in the blood of patients suspected of this diagnosis; a method for treating patients with acute myocardial infarction with antibody to marinobufagin, a bufodienolide, to prevent the occurrence of cardiac arrhythmias; antibodies which specifically recognize marinobufagin or other bufodienolides; hybridomas producing these antibodies; a process for preparing such antibodies; and an immunoassay method for marinobufagin for research purposes using its specific antibody.The antibodies of the present invention make it possible to conveniently measure bufodienolides with specificity and high sensitivity. This is useful in determining the existence and degree of hypertension and myocardial infarction, and in treating myocardial infarction.

Abstract: The present invention provides a novel method of determining the antagonist and agonist activity of a compound for steroid hormone receptors. The present invention also provides a method of determining antagonist activity of a compound for a hormone receptor by inducing the conformational change in the receptor. In addition, the present invention provides a novel method of determining the level of agonist activity of a compound for steroid hormone receptors.

Abstract: Corticotropin Releasing Factor-binding protein (CRF-BP) is produced recombinantly and is useful for modulating the biological activity of CRF. CRF-BP or fragments thereof and/or antibodies to such polypeptides are employed in diagnostic assays to determine the levels of CRF and CRF-BP and the ratio of CRF/CRF-BP in a vascular fluid sample. Following such an assay, pregnancy-related pathological disorders, such as increased risk of premature labor, can be treated, for example, by administering CRF-BP to lower the ratio of CRF/CRF-BP to within a normal range for pregnancy. Anti-CRF-BP antibodies are also useful to purify CRF-BP and to modulate the biological effect of CRF-BPs.

Abstract: The use of an antigen (or antibody) previously modified with a hapten makes the immunoassay of a trace component (analyte) on the basis of a change of turbidity or scattered light intensity caused by antigen-antibody reaction rapid and easy with high accuracy and high reproduction.

Abstract: Disclosed is a test cell and a method for detection of a preselected ligand in a liquid sample such as a body fluid. The test cell includes an elongate outer casing which houses an interior permeable material capable of transporting an aqueous solution and defining a sample inlet, a test volume, and a reservoir volume. The reservoir volume is disposed in a section of the test cell spaced apart from the inlet and is filled with sorbent material. The reservoir acts to receive liquid transported along a flow path defined by the permeable material and extending from the inlet and through the test volume. In the test volume is a test site which includes a first protein having a binding site specific to a first epitope of the ligand immobilized in fluid communication with the flow path. The test site can be observed through a window of the casing.

Abstract: Methods for determining the concentration of an analyte in a sample in which an analyte gradient is established and brought into contact with one or more zones that contain binding members that interact with the analyte and thereby produce a detectable signal. Devices that may be used to practice the disclosed methods are also provided.

Abstract: The present invention provides a new analytical method that an antigen, an antibody or a nucleic acid can be assayed at a high sensitivity in the immunoassay.The present invention provides a method for assaying a substance which comprises the steps of (a) reacting an antigen, an antibody or a nucleic acid which is the substance to be measured with an antibody, an antigen or a nucleic acid to which a polyacridinium compound is bound and which can be bound to the substance to be measured, (b) carrying out a luminescence treatment, and (c) measuring the luminescent intensity of the reaction solution.

Abstract: The present invention provides monoclonal antibodies that specifically recognize a specific structural portion of cardiac glycoside.The monoclonal antibodies of the present invention recognize an integrated structure in the steroid structure of cardiac glycoside or its aglycon comprising: a D-ring bonded in the cis-configuration with respect a C-ring; a methyl group bonded in the .beta.-configuration at the C13 position; a hydroxyl group bonded in the .beta.-configuration at the C14 position; and, an .alpha., .beta.-unsaturated lactone group bonded in the .beta.-configuration at the C17 position. This structure is common to cardiac glycoside substances that inhibit potassium ion concentration antagonist type Na.sup.+,K.sup.+ -ATPase.

Abstract: Immunoassay methods and reagents for the specific quantification of thyroxine in a test sample are disclosed employing antibodies prepared with thyroxine derivatives of the formula: ##STR1## wherein P is an immunogenic carrier material and X is a linking moiety. The present invention also describes the synthesis of unique labelled reagent of the formula: ##STR2## wherein Q is a detectable moiety and W is a linking moiety, preferably fluorescein or a fluorescein derivative.

Abstract: A device for performing an enzyme-labelled binding assay comprises an absorbent material and a developing solution containing a substrate for said enzyme, wherein the absorbent material is provided with a plurality of reagent zones including an indicator reagent zone and is capable of transporting the developing solution by capillary action sequentially through the reagent zones, and wherein the indicator reagent zone includes a reagent capable, directly or indirectly, of immobilising an enzyme-labelled reagent in an amount dependent on the assay result, characterized in that the absorbent material includes a reagent that prevents a signal formation except where enzyme-labelled reagent is immobilised at the indicator reagent zone. The absorbent material is suitably in the form of an elongate strip provided with transverse reagent zones. The device is useful for performing immunoassays, including immunometric assays and dual analyte assays.

Abstract: The present invention provides a method for predicting the outcome of IVF which comprises:(i) determining the level of 11.beta.-hydroxysteroid dehydrogenase (11.beta.-HSD) in a biological sample from a female patient; and(ii) predicting from the level of 11.beta.-HSD determined the probability of establishing pregnancy in said subject by IVF.It has been found that there is an inverse correlation between the levels of 11.beta.-HSD in the environment of the oocyte and subsequent establishment of pregnancy by IVF. The present invention also provides methods of screening female subjects for their suitability to take part in IVF programs, and kits to use in determining levels of 11.beta.-HSD.

Abstract: Methods for modulating the rates and dose responses of immunoassays through the incorporation of one or more detergents into the immunoassay reaction are disclosed. The methods are particularly suitable for automated immunoassay formats, especially with formats that use analyte-biotin bidentate reagents. The methods may be used to facilitate the detection of any desired, preselected pharmacological agent.

Abstract: An analytical test device useful for example in pregnancy testing, includes a hollow casing (500) constructed of moisture-impervious solid material, such as plastics materials, containing a dry porous carrier (510) which communicates indirectly with the exterior of the casing via a bibulous sample receiving member (506) which protrudes from the casing such that a liquid test sample can be applied to the receiving member and permeate therefrom to the porous carrier, the carrier containing in a first zone a labelled specific binding reagent is freely mobile within the porous carrier when in the moist state, and in a second zone spatially distinct from the first zone unlabelled specific binding reagent for the same analyte which unlabelled reagent is permanently immobilised on the carrier material and is therefore not mobile in the moist state, the two zones being arranged such that liquid sample applied to the porous carrier can permeate via the first zone into the second zone, and the device incorporating an a

Abstract: A diagnostic device is provided that determines the activity of CETP by the use of a new synthesized donor particle. The method in the diagnostic device for measuring the activity of cholesteryl ester transfer protein comprises: adding a prepared sonicated particle to a buffer to form a buffered solution, adding an Intralipid emulsion to the buffered solution for the purpose of accepting the transfer of neutral lipid, adding cholesteryl ester transfer protein to the buffered solution incubating the buffered solution, and reading the fluorescence of the buffer solution to measure the activity of the cholesteryl ester transfer protein. The synthesized donor particle is representative of a high density lipoprotein and comprises a fluorescent group, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino-covalently bound to a cholesteryl ester to form a NBD-CE core, a monolayer of phospholipid that surrounds the NBD-CE core and an apolipoprotein apoA-I associated with the monolayer and an aqueous phase.

Abstract: A method of determining a hapten which comprises: (1) contacting the sample which contains or may contain the hapten with a first binding partner which binds the hapten; (2) separating the hapten bound to the first binding partner from material which is not bound by the first binding partner; (3) contacting the hapten bound to the first binding partner with a second binding partner which binds the hapten; and (4) assaying the hapten bound to a binding partner as is described. Kits are also described.