Compositions and methods for assay measurement

Abstract

The present invention relates to novel compositions comprising an electrochemiluminescence (ECL) coreactant. In embodiments, the compositions further comprise an ionic component, a surfactant, or a combination thereof. In embodiments, the ECL coreactant is triethanolamine (TEA), tert-butyldiethanolamine (tBDEA), methyldibutylethanolamine (MDEA), 3-[bis-(2-hydroxyethyl)-amino]-propane-1-sulfonic acid (DEA-PS), or a combination thereof. Methods of using the compositions and kits containing the compositions are also provided herein.

Description

[Technical Field]

[0001] The present invention relates to compositions comprising electrochemiluminescent (ECL) co-reactants. In embodiments, the compositions further comprise ionic components, surfactants, or combinations thereof. In embodiments, the ECL co-reactants are triethanolamine (TEA), tert-butyldiethanolamine (tBDEA), methyldibutylethanolamine (MDEA), 3-[bis-(2-hydroxyethyl)-amino]-propane-1-sulfonic acid (DEA-PS), or combinations thereof. Methods of using the compositions and kits comprising the compositions are also provided. [Background technology]

[0002] Some commercially available instruments use electrochemiluminescence (ECL) for analytical measurements. Compounds that interact with ECL labels and produce ECL are called ECL co-reactants. A commonly used co-reactant is Ru(Bpy)3 +2Examples of ECL-derived tertiary amines for ECL (see, e.g., US5,846,485), oxalates, and persulfates, as well as hydrogen peroxide for ECL derived from luminol (see, e.g., US5,240,863). The light generated by ECL labeling can be used as a reporter signal in diagnostic procedures (see, e.g., US5,238,808). For example, the ECL label can be covalently bound to a detection reagent, and the involvement of the detection reagent in the binding interaction can be monitored by measuring the ECL released from the ECL label. Alternatively, the ECL signal from an ECL-active compound may indicate a chemical environment (see, e.g., US5,641,623, which describes an ECL assay for monitoring the formation or destruction of ECL co-reactants). ECL-based assays are available under US Patent Nos. 5,093,268, 5,147,806, 5,324,457, 5,591,581, 5,597,910, 5,641,623, 5,643,713, 5,679,519, 5,705,402, 5,846,485, 5,866,434, 5,786,141, 5,731,147, 6,066,448, and 6, Further details are provided in publications 136,268, 5,776,672, 5,308,754, 5,240,863, 6,207,369, 5,589,136, and 6,919,173, as well as in international publications 99 / 63347, 00 / 03233, 99 / 58962, 99 / 32662, 99 / 14599, 98 / 12539, 97 / 36931, and 98 / 57154.

[0003] Commercially available ECL instruments have become widely used due to their sensitivity, dynamic range, accuracy, and especially their tolerance for complex sample matrices. Several types of commercial instrumentation are available for performing ECL-based measurements (e.g., Debad, JD, et al., 2004. Clinical and Biological Applications of ECL, in: Electrogenerated Chemiluminescence. Marcel Dekker, pp. 43-78). ECL instruments are further described, for example, in U.S. Patents 5,935,779 and 5,993,740 (Bead-based ECL assay), U.S. Patents 6,140,045, 6,066,448, 6,090,545, and 6,207,369, and International Publication 98 / 12539 (ECL assay using immobilized binding reagent), U.S. Patents 6,977,722 and 7,842,246 (Multiwell plate with integrated electrode for ECL assay), and U.S. Publications 2012 / 0190589 and U.S.2012 / 0178091 (Cartridge-based ECL assay).

[0004] The ECL co-reactant tripropylamine (TPA) is typically used in ECL-based assays. [Overview of the Initiative]

[0005] In embodiments, the present invention provides a composition comprising (a) triethanolamine (TEA) and (b) an ionic component, wherein the composition has a pH of about 7.0 to about 8.0 and substantially contains no additional pH buffering components. In embodiments, the composition further comprises a surfactant.

[0006] In embodiments, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0 and substantially contains no additional pH buffering components.

[0007] In embodiments, the present invention provides a composition comprising (a) TEA, (b) an ionic component, and (c) optionally one or both of an ECL-labeled component and a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and optionally the composition substantially contains no additional pH buffering components.

[0008] In embodiments, the present invention provides a composition comprising (a) about 1000 mM to about 6500 mM triethanolamine (TEA) and (b) about 500 mM to about 2000 mM of an ionic component, wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the composition further comprises a surfactant. In embodiments, the composition further comprises an ECL-labeled component.

[0009] In embodiments, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) alkyl ether polyethylene glycol (PEG), wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the composition further comprises an ECL-labeled component.

[0010] In embodiments, the present invention provides a composition comprising (a) an electrochemiluminescent (ECL) co-reactant selected from N-tert-butyldiethanolamine (tBDEA), methyldiethanolamine (MDEA), 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid (DEA-PS), and combinations thereof; (b) an ionic component; and (c) a surfactant, wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the ECL co-reactant is tBDEA. In embodiments, the ECL co-reactant is MDEA. In embodiments, the ECL co-reactant is DEA-PS, and the composition further comprises an ECL-labeled component.

[0011] In embodiments, the present invention provides a method for generating electrochemiluminescence (ECL), comprising: (a) contacting an electrode with (i) a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant, and (ii) an ECL label; and (b) applying a voltage to the electrode to generate ECL.

[0012] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) contacting a liquid sample with a surface, wherein the surface comprises a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant, and the liquid sample comprises an ECL-labeled component, or the liquid sample comprises a binding partner of an ECL-labeled component, and the method further comprises contacting the surface with an ECL-labeled component to thereby form a bound complex comprising an ECL-labeled component on the surface; (b) applying a voltage to the surface to generate ECL; and (c) detecting the generated ECL to thereby detect a bound complex.

[0013] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) contacting a liquid sample with a surface, the surface comprising an ECL-labeled component and a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant, wherein the liquid sample comprises a binding partner of the ECL-labeled component, thereby forming a bound complex comprising the ECL-labeled component on the surface; (b) applying a voltage to the surface to generate ECL; and (c) detecting the generated ECL, thereby detecting the bound complex.

[0014] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) forming a bound complex on a surface, wherein the bound complex contains an ECL-labeling component; (b) contacting the bound complex with a composition provided herein, or a TEA composition containing TEA, an ionic component, and optionally a surfactant; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex.

[0015] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) forming a bound complex on a surface, wherein the surface optionally includes an electrode, and the bound complex comprises a binding reagent immobilized on the surface and a detection reagent including an electrochemiluminescence (ECL) label; (b) contacting the bound complex with a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex.

[0016] In embodiments, the present invention provides a method for detecting a target analyte in a sample, comprising: (a) contacting the sample with a surface containing a binding reagent, the binding reagent being specifically bound to the analyte; and (ii) a detection reagent being specifically bound to the analyte, the detection reagent comprising an electrochemiluminescence (ECL) label, thereby forming a binding complex on the surface comprising the binding reagent, the analyte, and the detection reagent; (b) contacting the binding complex on the surface with a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL.

[0017] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) forming an assay mixture by combining a sample with a detection mixture comprising (i) a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant, and (ii) at least two copies of a detection reagent, each copy of the detection reagent comprising an ECL label; (b) contacting the assay mixture with a bound reagent immobilized on a surface comprising an electrode, under conditions that (I) a bound complex is formed on the surface, the bound complex comprises the bound reagent and a first copy of the detection reagent, and (II) a second copy of the detection reagent remains in the solution; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex.

[0018] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) an assay mixture comprising (i) a bound reagent immobilized on a surface, the surface optionally comprising an electrode; and (ii) a detection mixture comprising at least two copies of a detection reagent, each copy of the detection reagent comprising an electrochemiluminescence (ECL) label; (i) incubation under conditions such that a bound complex is formed on the surface, the bound complex comprises the bound reagent and a first copy of the detection reagent, and (ii) a second copy of the detection reagent remains in solution; (b) contacting the bound complex with a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex.

[0019] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) an assay mixture comprising (i) a bound reagent immobilized on a surface, the surface optionally comprising an electrode; (ii) a detection mixture comprising at least two copies of a detection reagent, each copy of the detection reagent comprising an electrochemiluminescence (ECL) label; and (iii) a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant; (i) incubation under conditions that a bound complex is formed on the surface, the bound complex comprising the bound reagent and a first copy of the detection reagent, and (ii) a second copy of the detection reagent remaining in solution; (b) applying a voltage to the surface to generate ECL; and (c) detecting the generated ECL to detect the bound complex.

[0020] In embodiments, the present invention provides a method for generating electrochemiluminescence (ECL), comprising: (a) contacting an electrode with (i) a composition provided herein and (ii) an ECL label; (b) applying a voltage to the electrode; and (c) generating ECL.

[0021] In embodiments, the present invention provides a method for quantifying the amount of electrochemiluminescence (ECL) label in a sample, comprising: (a) contacting an electrode with (i) a composition provided herein and (ii) the sample containing the ECL label; (b) applying a voltage to the electrode; (c) generating ECL; (d) measuring the ECL; and (e) quantifying the amount of ECL label from the measured ECL.

[0022] In embodiments, the present invention provides a method for producing a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) a surfactant, wherein the method does not involve the addition of an additional pH buffering component.

[0023] In an embodiment, the present invention provides an assay module comprising a dry TEA composition, wherein the TEA composition comprises TEA, an ionic component, and optionally a surfactant.

[0024] In embodiments, the present invention provides a kit comprising (a) a composition provided herein, or a TEA composition comprising TEA, an ionic component, and optionally a surfactant, and (b) optionally a surface comprising an electrode, wherein the TEA composition optionally does not contain additional pH buffering components.

[0025] In embodiments, the present invention provides a kit comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) a surfactant in one or more containers, vials, or compartments, wherein the kit does not contain additional pH buffering components. [Brief explanation of the drawing]

[0026] The following drawings form part of this specification and are included to further illustrate exemplary embodiments of particular aspects of the invention.

[0027] [Figure 1A] In relation to Example 1, the results of one embodiment of an ECL-based assay are shown. A panel of ECL co-reactants combined with one of two surfactants was tested in a solid surface ECL assay for ECL generation and its ability to distinguish between surface-bound ("BTI") ECL labels and free (in solution, "FT") ECL labels. Figure 1A shows the ECL signals measured using only ECL reading buffer (without labeling) for BTI, FT, and background signal ("D100"). [Figure 1B] Figure 1B shows the ratio of the ECL signal from the coupled label to the ECL signal from the free label ("BTI / FT") and the signal-to-background ratio ("S / B"). [Figure 2A]In relation to Example 2, the results of one embodiment of the ECL-based assay are shown. Figure 2A shows plots of ECL generated from BTI and FT at different TEA concentrations, as well as the BTI / FT ratio. [Figure 2B] Figure 2B shows the measured ECL signals from BTI, FT, and background (D100) at different TEA concentrations. [Figure 2C] Figure 2C shows the BTI / FT ratio, S / B ratio, and ECL generation percentage compared to the PIPES ECL read buffer. [Figure 3A] In relation to Example 3, the results of one embodiment of the ECL-based assay are shown. Figure 3A shows the change in the ECL signal as a function of PIPES concentration. [Figure 3B] Figure 3B shows the change in the ECL signal as a function of PIPES or TEA concentration. [Figure 4A] This specification describes embodiments of the ECL-based assay, further described in Example 4. Figure 4A shows a "standard" two-step washing assay in which a capture antibody ("cAb", conjugation reagent) immobilized on a surface is brought into contact with a mixture of analytes (one of which is specifically bound to the capture antibody), and the surface is then washed, resulting in the capture of the analytes on the surface. Next, a mixture of detection antibodies ("dAb", detection reagent), each containing an ECL label and one of which is specifically bound to the analyte, is added to the surface, and the surface is then washed, resulting in a conjugation complex containing cAb, the analyte, and dAb. Next, an ECL reading buffer is added to the surface, and the resulting ECL is read by an ECL reading instrument. [Figure 4B] Figure 4B shows a "one-step" assay in which the captured antibody on the surface is brought into contact with the analyte mixture, and then the surface is washed as shown in Figure 4A. Next, the detection antibody mixture is added, followed by the ECL reading buffer, but no washing is performed between the addition of the detection antibody mixture and the ECL reading buffer. Then, the generated ECL is read by an ECL reading instrument. [Figure 4C]Figure 4C shows a "one-step no-wash" assay in which the surface-borne capture antibody is brought into contact with the analyte mixture and the detection antibody mixture, and then into contact with the ECL reading buffer, without washing between any of the steps. The resulting ECL is then read by an ECL reader. [Figure 4D] Figure 4D shows a "mock ECL-labeled" assay in which a captured antibody on a surface is brought into contact with the analyte mixture, the surface is washed, the detection antibody mixture is added, and the surface is optionally washed again, resulting in a conjugated complex as shown in Figure 4A. Next, an ECL reading buffer is added to the surface along with a detection antibody that contains the ECL label but does not bind to any component of the conjugated complex on the surface, which acts as a proxy for the "free" ECL label in solution. The resulting ECL is then read by an ECL reading instrument. [Figure 4E] Figure 4E shows a multiplexed version of a "standard" two-step washing assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody capable of binding to an analyte in the analyte mixture. Washing the surfaces containing the binding domains after adding the analyte mixture results in multiple analytes being captured on the binding domains. Next, a mixture of detection antibodies, each containing an ECL label and capable of binding to an analyte in the analyte mixture, is added to the surface, and the surface is then washed, resulting in multiple binding complexes, each binding complex containing a cAb, an analyte, and a dAb. Next, ECL reading buffer is added to the surface, and the resulting ECL is read by an ECL instrument. [Figure 4F] Figure 4F shows a multiplexed version of the "one-step" assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody capable of binding to an analyte in the analyte mixture. The surfaces containing the binding domains are washed after adding the analyte mixture as shown in Figure 4E. The detection antibody mixture is added to form multiple binding complexes, and then the ECL reading buffer is added, but without washing between the addition of the detection antibody mixture and the ECL reading buffer. The resulting ECL is then read by an ECL reading instrument. [Figure 4G] Figure 4G shows a multiplexed version of a "one-step no-wash" assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody capable of binding to an analyte in the analyte mixture. The surfaces containing the binding domains are brought into contact with the analyte mixture and the detection antibody mixture to form multiple binding complexes, and then ECL reading buffer is added, without washing between any steps. The resulting ECL is then read by an ECL reading instrument. [Figure 4H] Figure 4H shows a multiplexed version of the “mock ECL-labeled” assay, in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody capable of binding to an analyte in the analyte mixture. The surfaces containing the binding reagents are brought into contact with the analyte mixture, the surfaces are washed, the detection antibody mixture is added, and the surfaces are optionally washed again, resulting in multiple binding complexes as shown in Figure 4E. Next, ECL reading buffer is added to the surfaces along with a detection antibody containing the ECL label but not binding to any component of the binding complexes on the surface, which acts as a proxy for the “free” ECL label in solution. The resulting ECL is then read by an ECL reading instrument. [Figure 5A] In relation to Example 5A, the results of one embodiment of an ECL-based assay are shown. Figure 5A shows the results of specific ECL signals and nonspecific binding (NSB) from three different multiplexed assay formats (shown in Figures 4E, 4F, and 4H) using BDEA, PIPES, and TEA reading buffers. [Figure 5B] Figure 5B shows the limit of detection (LLOD) of the assay in Figure 5A. [Figure 5C] Figure 5C shows a relative comparison of the ECL and NSB results from Figure 5A. [Figure 5D] Figure 5D shows a comparison of signal-to-background (S / B) and signal-to-noise (S / N) ratios across all ECL read buffers and assay formats. [Figure 6A]In relation to Example 5B, the results of one embodiment of an ECL-based assay are shown. Figure 6A shows the results of specific ECL signals and nonspecific binding (NSB) from three different multiplexed assay formats (shown in Figures 4E, 4F, and 4G) using BDEA, PIPES, and TEA reading buffers. [Figure 6B] Figure 6B shows the limit of detection (LLOD) of the assay in Figure 6A. [Figure 6C] Figure 6C shows a relative comparison of the ECL and NSB results from Figure 6A. [Figure 7A] In relation to Example 6, the results of one embodiment of the ECL-based assay are shown. Figure 7A shows a list of sample matrices tested using TEA reading buffer in a one-step, no-wash ECL assay. [Figure 7B] Figure 7B shows a list of interfering substances added to the sample matrix shown in Figure 7A, which are tested using TEA reading buffer in a one-step, no-wash ECL assay. [Figure 8A] Figure 8A shows the results of ECL signals generated from TEA reading buffers containing bound ECL labeling ("bound") and free ECL labeling ("free") using different sample matrices mixed with diluents. "H2O" indicates the signal from a control where water was used instead of the sample matrix before the TEA reading buffer. The column header containing "free" indicates 6 nM of free ECL labeling in the diluent. [Figure 8B] Figure 8B shows the results of Figure 8A, normalized for the ECL signal generated from an assay without the sample matrix. [Figure 9A] Figure 9A shows the results of ECL signals generated from TEA reading buffers containing bound ECL labels and free ECL labels with different interfering substances in different sample matrices. [Figure 9B] Figure 9B shows the results of Figure 9A, normalized for the ECL signal generated from assays without the addition of sample matrix and interfering substances. [Figure 10A]Figure 10A shows the results of ECL signals generated from TEA reading buffers containing free ECL labeling ("D3+STAG") in different sample matrices. The column header containing "free" indicates 240 nM of free ECL labeling in the diluent. [Figure 10B] Figure 10B shows the results of Figure 10A, normalized for the ECL signal generated from an assay without sample matrix addition. [Figure 11A] Figure 11A shows the results of ECL signals generated from TEA reading buffers containing 240 nM free ECL with different interfering substances in different sample matrices. [Figure 11B] Figure 11B shows the results of Figure 11A, normalized for the ECL signal generated from assays without the addition of sample matrix and interfering substances. [Figure 12] In relation to Example 7, the results of one embodiment of an ECL-based assay are shown. The combination of ECL co-reactants described in Example 1 was tested in an ECL-based assay. The upper right side of the chart in Figure 12 shows the ECL signal generated from BTI, and the lower left side of the chart in Figure 12 shows the ratio of the ECL signal of the mixed ECL co-reactants to the sum of the signals generated by the individual ECL co-reactants. [Figure 13A] In relation to Example 8, the results of one embodiment of an ECL-based assay are shown. The ECL co-reactants described in Example 1 were tested for their sensitivity to the presence of TRITON® X-100. Figure 13A shows the ECL signals from BTI and FT for each ECL reactant in TRITON® X-100 (TX100) and PEG(18) tridecyl ether (PEG18TDE). [Figure 13B] Figure 13B shows the ratio of ECL produced in TRITON® X-100 to PEG(18) tridecyl ether. [Modes for carrying out the invention]

[0028] The ECL co-reactants of the present invention provide consistent ECL production across different assay formats. For example, the compositions of this specification comprising triethanolamine (TEA) have been found useful in ECL-based assays that do not require a washing step. Many ECL-based assays performed on solid surfaces include at least one washing step to remove unbound ECL labels before detecting ECL labels on the surface (i.e., "washing" assays). The washing step may be eliminated if the detection method can effectively distinguish between ECL labels bound to the surface (e.g., as part of the binding complex being detected) or unbound "free" ECL labels in solution. "No-wash" assay formats, which eliminate the washing step, are often advantageous because performing the washing step can be difficult or cumbersome in many situations. However, no-wash assay formats are typically difficult to develop due to high background ECL signals resulting from incomplete distinction between free and bound ECL labels present in the reaction mixture.

[0029] In ECL-based assays performed on solid surfaces, triethanolamine (TEA) has been found to remarkably distinguish, to a high degree, between unbound ("free") ECL labels in solution and surface-bound ECL labels. The compositions described herein, comprising TEA, increase the ratio of ECL signals from bound labels to ECL signals from free labels compared to conventional compositions containing conventional co-reactants such as tripropylamine (TPA). Thus, the compositions herein offer improved assay performance, particularly when measuring low-affinity interactions, which require the presence of high concentrations of ECL labels in the reaction, but which are also expected to suffer significant signal loss due to the dissociation of bound complexes during the washing step.

[0030] For example, the ECL signals generated from compositions herein, comprising TEA, tert-butyldiethanolamine (tBDEA), methyldiethanolamine (MDEA), and / or 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid (DEA-PS), offer further advantages, such as improved consistency of performance between different compositions based on the presence or absence of surfactants, or based on the identity of the surfactants. In particular, compositions function similarly whether they contain no surfactant, contain a mild surfactant that does not disrupt the lipid bilayer membrane (such as polyethylene glycol (18) tridecyl ether), or contain a more severe surfactant (such as TRITON® X-100). Therefore, a severe surfactant (e.g., TRITON® X-100, which disrupts the lipid bilayer membrane that is part of a specific target analyte such as whole cells or extracellular vesicles) is not required in compositions containing ECL co-reactants described herein, in contrast to tripropylamine (TPA), a typical ECL co-reactant that usually requires TRITON® X-100 for optimal ECL generation. Therefore, the compositions herein are useful in assays for detecting analytes sensitive to strict surfactants. Furthermore, the ECL signals of ECL co-reactants are not significantly affected by the presence of different surfactants, and thus these ECL co-reactants are versatile and can be easily incorporated into different formulations while maintaining their ECL-generating ability.

[0031] Therefore, the compositions of this specification, for example, comprising TEA, tBDEA, MDEA, and / or DEA-PS, advantageously expand the types of viable ECL-based assays.

[0032] Unless otherwise defined herein, scientific and technical terms used in this disclosure shall have meanings generally understood by those skilled in the art. Furthermore, unless otherwise required by context, singular terms shall include plural forms and plural terms shall include singular forms. The articles “a” and “an” are used herein to refer to one or more (i.e., at least one) grammatical objects of the article. For example, “an element” means one or more elements.

[0033] The use of the term “or” in the claims is used to mean “and / or” unless it is explicitly indicated that it refers only to substitutes, or substitutes are mutually exclusive, but this disclosure supports the definition that refers only to substitutes and “and / or”.

[0034] As used herein, the terms “comprising” (and any variations or forms of “comprising,” such as “comprise” and “comprises”), “having” (and any variations or forms of “having,” such as “have” and “has”), “including” (and any variations or forms of “including,” such as “includes” and “include”), or “containing” (and any variations or forms of “containing,” such as “contains” and “contain”) are either comprehensive or open-ended and do not exclude any additional elements or method steps not described herein.

[0035] The use of the term "for example" and its corresponding abbreviation "eg" (whether it is an italicized pair or not) means that the particular terms described are representative examples and embodiments of the invention, and are not intended to be limited to the specific examples referenced or cited, unless expressly otherwise specified.

[0036] As used herein, “between” is a range that includes both ends of the range. For example, the numbers between x and y explicitly include the numbers x and y, and any numbers (including fractions and integers) contained within x and y. Furthermore, references to the range “5 to 10” herein include the integers 5, 6, 7, 8, 9, and 10, as well as fractions such as 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, and 5.9. References to any numerical range explicitly include each numerical value (including fractions and integers) that the range encompasses. For example, the range “at least 50” or “at least about 50” includes integers such as 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, and 60, and fractions such as 50.1, 50.2, 50.3, 50.4, 50.5, 50.6, 50.7, 50.8, and 50.9. In further examples, references to the range “less than 50” or “less than about 50” in this specification include integers such as 49, 48, 47, 46, 45, 44, 43, 42, 41, and 40, and fractions such as 49.9, 49.8, 49.7, 49.6, 49.5, 49.4, 49.3, 49.2, 49.1, and 49.0.

[0037] As used herein, the terms “substantially” or “substantial” are applicable when used in a negative sense to indicate the complete or near-complete absence of an action, feature, characteristic, state, structure, item, or result. For example, a “substantially” flat surface would either be perfectly flat or nearly flat with the same effect as being perfectly flat. In further embodiments, a composition “substantially” free of a particular component would either have no amount of that component at all, or the component would be present in the composition in such a low amount that it would have the same effect as if the component were not present.

[0038] In embodiments, the present invention provides a composition comprising (a) triethanolamine (TEA) and (b) an ionic component, wherein the composition has a pH of about 7.0 to about 8.0 and substantially contains no additional pH buffering components.

[0039] In embodiments, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0 and substantially contains no additional pH buffering components.

[0040] In embodiments, the present invention provides a method for a composition comprising (a) about 1000 mM to about 6500 mM triethanolamine (TEA) and (b) about 500 mM to about 2000 mM of an ionic component, wherein the composition has a pH of about 7.0 to about 8.0.

[0041] In embodiments, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) alkyl ether polyethylene glycol (PEG), wherein the composition has a pH of about 7.0 to about 8.0.

[0042] In embodiments, the present invention provides a composition comprising (a) TEA, (b) an ionic component, and (c) optionally one or both of an ECL-labeled component and a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and optionally the composition substantially contains no additional pH buffering components.

[0043] In embodiments, the present invention provides a composition comprising (a) an electrochemiluminescent (ECL) co-reactant selected from N-tert-butyldiethanolamine (tBDEA), methyldiethanolamine (MDEA), 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid (DEA-PS), and combinations thereof; (b) an ionic component; and (c) a surfactant, wherein the composition has a pH of about 7.0 to about 8.0.

[0044] In embodiments, the present invention provides compositions comprising, or essentially comprising, the described components in described amounts. In compositions essentially comprising the described components, such compositions specifically exclude components that substantially (materially) affect the ECL-generating properties of the composition. The ECL-generating properties of the composition can be determined by methods known to those skilled in the art. For example, the composition can be brought into contact with a known amount of ECL label on an electrode, and a voltage can be applied to the electrode to generate ECL. In embodiments, “substantially (materially) unaffected” ECL-generating properties mean that a composition “essentially comprising” the described components generates about 80%, about 90%, about 95%, about 98%, about 99%, about 100%, about 101%, about 102%, about 105%, about 110%, or about 120% ECL as a composition “comprising” the described components. In embodiments, compositions essentially comprising the described components specifically exclude additional ECL-generating compounds, such as additional ECL co-reactants.

[0045] In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) a surfactant, wherein the composition has a pH of about 7.0 to about 8.0 and is substantially free of additional pH buffering components. In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) a surfactant, wherein the composition has a pH of about 7.0 to about 8.0 and is substantially free of additional pH buffering components.

[0046] In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, (c) a surfactant, and (d) an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0 and substantially contains no additional pH buffering components. In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, (c) a surfactant, and (d) an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0.

[0047] In an embodiment, the present invention provides a composition comprising (a) about 1000 mM to about 6500 mM triethanolamine (TEA), (b) about 500 mM to about 2000 mM ionic components, and (c) a surfactant, wherein the composition has a pH of about 7.0 to about 8.0. In an embodiment, the present invention provides a composition comprising (a) about 1000 mM to about 6500 mM triethanolamine (TEA), (b) about 500 mM to about 2000 mM ionic components, and (c) a surfactant, wherein the composition has a pH of about 7.0 to about 8.0.

[0048] In an embodiment, the present invention provides a composition comprising (a) about 1000 mM to about 6500 mM triethanolamine (TEA), (b) about 500 mM to about 2000 mM ionic component, (c) surfactant, and (d) ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0. In an embodiment, the present invention provides a composition comprising (a) about 1000 mM to about 6500 mM triethanolamine (TEA), (b) about 500 mM to about 2000 mM ionic component, (c) surfactant, and (d) ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0.

[0049] In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) alkyl ether polyethylene glycol (PEG), wherein the composition has a pH of about 7.0 to about 8.0. In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) alkyl ether polyethylene glycol (PEG), wherein the composition has a pH of about 7.0 to about 8.0.

[0050] In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, (c) an alkyl ether-PEG, and (d) an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0. In an embodiment, the present invention provides a composition comprising (a) triethanolamine (TEA), (b) an ionic component, (c) an alkyl ether-PEG, and (d) an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0.

[0051] In embodiments, the present invention provides a composition comprising (a) an electrochemiluminescent (ECL) co-reactant selected from N-tert-butyldiethanolamine (tBDEA), methyldiethanolamine (MDEA), 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid (DEA-PS), and combinations thereof; (b) an ionic component; (c) a surfactant; and (d) a pH interfering component, wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the present invention provides a composition comprising (a) an electrochemiluminescent (ECL) co-reactant selected from N-tert-butyldiethanolamine (tBDEA), methyldiethanolamine (MDEA), 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid (DEA-PS), and combinations thereof; (b) an ionic component; (c) a surfactant; and (d) a pH interfering component, wherein the composition has a pH of about 7.0 to about 8.0.

[0052] In an embodiment, the present invention provides a composition comprising (a) an ECL co-reactant selected from tBDEA, MDEA, DEA-PS, and combinations thereof, (b) an ionic component, (c) a surfactant, (d) a pH buffering component, and (e) an ECL labeling component, wherein the composition has a pH of about 7.0 to about 8.0. In an embodiment, the present invention provides a composition comprising (a) an ECL co-reactant selected from tBDEA, MDEA, DEA-PS, and combinations thereof, (b) an ionic component, (c) a surfactant, (d) a pH buffering component, and (e) an ECL labeling component, wherein the composition has a pH of about 7.0 to about 8.0.

[0053] As discussed herein, the ECL co-reactants herein are advantageous in combination with different classes of surfactants (e.g., mild surfactants that do not disrupt the lipid bilayer, and more severe surfactants that can disrupt the lipid bilayer) across different assay formats (e.g., wash and non-wash assays) to provide consistent ECL production. Accordingly, compositions comprising the ECL co-reactants herein (also referred to as “ECL reading buffers”) are useful in a wide range of ECL-based binding assays.

[0054] In the embodiment, the ECL co-reactant includes a tertiary amine. In the embodiment, the ECL co-reactant includes a tertiary alkylamine. In the embodiment, the ECL co-reactant includes a tertiary hydroxyalkylamine. In the embodiment, the ECL co-reactant includes an amphoteric tertiary amine. In the embodiment, the ECL co-reactant includes a secondary amine. In the embodiment, the ECL co-reactant is tributylamine (TBA), (dibutyl)aminoethanol (DBAE), (diethyl)aminoethanol (DEAE), triethanolamine (TEA), butyldiethanolamine (BDEA), propyldiethanolamine (PDEA), ethyldiethanolamine (EDEA), methyldiethanolamine (MDEA), tert-butyldiethanolamine (tBDEA), dibutylamine (DBA), butylethanolamine (BEA), diethanolamine (DEA), dibutylaminepropylsulfonate (DBA-PS), dibutylaminebutylsulfonate (DBA-BS), butylethanolaminepropylsulfonate (BEA-PS), butylethanolaminebutylsulfonate (BEA-BS), diethanolaminepropylsulfonate (also known as 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid, DEA-PS), or diethanolaminebutylsulfonate (DEA-BS). The structures of exemplary ECL co-reactants described herein are shown below. [ka]

[0055] Triethanolamine ECL co-reactant The present invention provides compositions comprising an ECL co-reactant. In embodiments, the ECL co-reactant is triethanolamine (TEA). As discussed herein, TEA has been found to provide advantageous ECL generation properties in no-wash binding assays. In assays in which a target species (e.g., an analyte or binding complex described herein) is captured and detected on a solid surface, TEA can distinguish between "free" ECL labels (e.g., analyte or binding complex) that are not part of the species being detected and "bound" ECL labels (e.g., analyte or binding complex) that are part of the species bound to the surface. No-wash binding assays utilizing TEA as an ECL co-reactant reduce non-specific ECL from the ECL label relative to the detected species, thereby reducing background ECL and increasing the signal-to-background ratio of the assay. In the embodiments, a no-wash assay using TEA as the ECL co-reactant has a signal-to-background ratio that is 2 times, 3 times, 4 times, 5 times, or 10 times higher than assays using tripropylamine (TPA) or piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES) as the ECL co-reactant. In the embodiments, a no-wash assay using TEA as the ECL co-reactant has a detection limit that is 2 times, 3 times, 4 times, 5 times, 10 times, 20 times, or 40 times lower than assays using TPA or PIPES as the ECL co-reactant.

[0056] A further advantage of TEA as an ECL co-reactant is that TEA is insensitive to sample matrix and / or interfering substances. This is particularly beneficial in connection with no-wash assays where the reaction mixture may contain a matrix of human or animal source (e.g., containing proteins, cellular components and debris, culture media, etc.), which may also contain metabolites and / or drug interfering substances such as acetaminophen, ibuprofen, naproxen, salicylic acid, and / or tolbutamine. In embodiments, TEA produces substantially the same ECL signal in a reaction mixture containing one or more sample matrices and / or one or more interfering substances as it does in a reaction mixture without sample matrix and / or interfering substances.

[0057] It has been further discovered that TEA offers the advantage of producing a consistent ECL signal when used in the absence of surfactants, or when combined with different types of surfactants, such as the strict and mild surfactants described herein. When used herein, “strict” surfactants can disrupt, lyse, and / or dissolve lipid bilayer membranes (e.g., membranes of cells or extracellular vesicles (EVs)). In contrast, “mild” surfactants do not disrupt, lyse, or dissolve lipid bilayer membranes. In embodiments, compositions comprising TEA and a strict surfactant produce substantially similar ECL signals as compositions containing the same components, except that a mild surfactant is present instead of a strict surfactant, when subjected to the same ECL generation conditions (e.g., voltage waveform, electrode type, amount of composition, amount of ECL label, etc.). In the embodiments, the strict surfactants are TRITON® X-100, TRITON® X-114, NP-40, IGEPAL® CA-630, 3-[(3-colamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), or sodium dodecyl sulfate (SDS). In the embodiments, the mild surfactants are BRIJ®, TWEEN®, PLURONIC®, or KOLLIPHOR® surfactants, or alkyl ether-PEG surfactants such as PEG(18) tridecyl ether.

[0058] TEA has a pKa of about 7.7 and is capable of maintaining the pH of the composition within the range of about 7.0 to about 8.0, which is a typical desired pH range for biological assays. Furthermore, TEA compositions with a pH of about 7.0 to about 8.0 have an ECL signal that is preferentially produced from electrode-bound ECL labels compared to unbound ECL labels described herein. Thus, the compositions herein comprising TEA have the additional advantage of pH compatibility with biological assays and do not require additional pH buffering components, thereby simplifying the composition production process and reducing costs. In embodiments, compositions comprising TEA substantially contain no additional pH buffering components. Materials that can act as pH buffering components to maintain a solution within a specific pH range are known to those skilled in the art. For example, buffers that can maintain a pH of approximately 7.0 to 8.0 include piperazine N,N'-bis(2-ethanesulfonic acid) (PIPES), cholamine chloride, 3-(N-morpholino)propanesulfonic acid (MOPS), N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid (TES), 3-(N,N-bis[2-hydroxyethyl]amino)-2-hydroxypropanesulfonic acid (DIPSO), 4-(N-morpholino)butanesulfonic acid (MOBS), acetamidoglycine, and N-[tris(hydroxymethyl)methyl] Examples of pH buffering components include, but are not limited to, tris(hydroxymethyl)aminomethane ("Tris"), piperazine-1,4-bis(2-hydroxypropanesulfonic acid) dihydrate (POPSO), N-(hydroxyethyl)piperazine-N'-2-hydroxypropanesulfonic acid (HEPPSO), 3-[4-(2-hydroxyethyl)piperazine-1-yl]propane-1-sulfonic acid (HEPPS), tricine, glycinamide, N-(2-hydroxyethyl)piperazine-N'-(4-butanesulfonic acid) (HEPBS), and bicine. Further non-limiting examples of pH buffering components include tris(hydroxymethyl)aminomethane ("Tris"), phosphates, HEPES, glycylglycine ("GlyGly"), borates, acetates, and citrates.In the embodiments, the composition containing TEA does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, or citrates. In the embodiments, the composition containing TEA is substantially free of additional components having a pKa of about 7.0 to about 8.0.

[0059] Furthermore, many common pH buffering compounds have tertiary amines in their structure and can generate ECL. Exemplary pH buffering compounds that can act as ECL co-reactants are provided in US6,919,173 and include, but are not limited to, HEPES, POPSO, HEPPSO, and PIPES. In embodiments, compositions containing TEA substantially do not contain additional ECL co-reactants. In embodiments, compositions containing TEA do not contain any of HEPES, POPSO, HEPPSO, and PIPES. In embodiments, compositions containing TEA do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES.

[0060] In the embodiment, the concentration of TEA in the composition is approximately 500 mM to approximately 7000 mM, approximately 800 mM to approximately 6800 mM, approximately 1000 mM to approximately 6500 mM, approximately 1000 mM to approximately 6400 mM, approximately 1000 mM to approximately 6000 mM, approximately 1000 mM to approximately 5500 mM, approximately 1100 mM to approximately 5000 mM, and approximately The molecular weight ranges are approximately 1100mM to 4800mM, approximately 1100mM to 4000mM, approximately 1100mM to 3500mM, approximately 1100mM to 3200mM, approximately 1100mM to 3000mM, approximately 1100mM to 2500mM, approximately 1200mM to 2400mM, or approximately 1200mM to 1600mM. In the embodiment, the concentration of TEA in the composition is approximately 1000 mM, approximately 1100 mM, approximately 1200 mM, approximately 1300 mM, approximately 1400 mM, approximately 1500 mM, approximately 1600 mM, approximately 1700 mM, approximately 1800 mM, approximately 1900 mM, approximately 2000 mM, approximately 2100 mM, approximately 2200 mM, approximately 230 mM 0mM, approx. 2400mM, approx. 2500mM, approx. 2600mM, approx. 2700mM, approx. 2800mM, approx. 2900mM, approx. 3000mM, approx. 310 0mM, approx. 3200mM, approx. 3300mM, approx. 3400mM, approx. 3500mM, approx. 3600mM, approx. 3700mM, approx. 3800mM, approx. 390 0mM, approx. 4000mM, approx. 4100mM, approx. 4200mM, approx. 4300mM, approx. 4400mM, approx. 4500mM, approx. 4600mM, approx. 4700mM, approx. 4800mM, approx. 4900mM, approx. 5000mM, approx. 5100mM, approx. 5200mM, approx. 5300mM, approx. 5400mM, approx. 550 The values ​​are 0 mM, approximately 5600 mM, approximately 5700 mM, approximately 5800 mM, approximately 5900 mM, approximately 6000 mM, approximately 6100 mM, approximately 6200 mM, approximately 6300 mM, approximately 6400 mM, approximately 6500 mM, approximately 6600 mM, approximately 6700 mM, approximately 6800 mM, approximately 6900 mM, or approximately 7000 mM. In one embodiment, the concentration of TEA in the composition is at least about 1000 mM, at least about 1200 mM, at least about 1600 mM, at least about 1800 mM, at least about 2000 mM, at least about 2500 mM, at least about 3000 mM, at least about 3500 mM, at least about 4000 mM, at least about 4500 mM, at least about 5000 mM, at least about 5500 mM, or at least about 6000 mM.Surprisingly, the TEA concentration in the composition showed a positive correlation with the intensity of the generated ECL signal, which was unexpected because other ECL co-reactants such as PIPES (both PIPES and TEA were expected to behave similarly to TEA, as both have the ability to confine ECL near the electrode as described herein) showed a decrease in ECL generation with increasing ECL co-reactant concentrations (see, for example, Figures 3A and 3B). Therefore, the TEA compositions provided herein can preferentially and consistently generate ECL signals from electrode-bound ECL labels over a wide concentration range, e.g., about 1000 mM to about 6500 mM, compared to unbound ECL labels described herein. The consistency of electrode-bound ECL generation reduces variability in ECL generation in ECL-based assays, for example, in wash or non-wash assay formats. ECL co-reactants that can be used at high concentrations (e.g., TEA) offer advantages in no-wash assays by minimizing the dilution of the sample and / or assay mixture, avoiding disturbances to the binding equilibrium and dynamics of assay components, and thus maximizing the ECL signal. ECL co-reactants that can be used at high concentrations (e.g., TEA) are also useful in assays with lower affinity binding and / or detection reagents, providing improved sensitivity compared to ECL co-reactants that cannot be used at high concentrations (e.g., PIPES).

[0061] Alkyldiethanolamine / amphoterionic tertiary amine ECL co-reactant The present invention further provides compositions comprising alkyldiethanolamine ECL co-reactants and / or amphoteric tertiary amine ECL co-reactants. In embodiments, the alkyldiethanolamine is butyldiethanolamine (BDEA), propyldiethanolamine (PDEA), ethyldiethanolamine (EDEA), methyldiethanolamine (MDEA), or tert-butyldiethanolamine (tBDEA). In embodiments, the alkyldiethanolamine is N-tert-butyldiethanolamine (tBDEA) or methyldiethanolamine (MDEA). In embodiments, the amphoteric tertiary amine ECL co-reactant is dibutylamine propyl sulfonate (DBA-PS), dibutylamine butyl sulfonate (DBA-BS), butylethanolamine propyl sulfonate (BEA-PS), butylethanolamine butyl sulfonate (BEA-BS), diethanolamine propyl sulfonate (also known as 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid, DEA-PS), or diethanolamine butyl sulfonate (DEA-BS). In embodiments, the amphoteric tertiary amine ECL co-reactant is DEA-PS. tBDEA, MDEA, and DEA-PS have been found to exhibit advantageous consistent ECL formation properties when used in the absence of surfactants or in combination with different types of surfactants, such as the abrasive and mild surfactants described herein. In the embodiments, compositions comprising tBDEA, MDEA, and / or DEA-PS, and a strict surfactant, when subjected to the same ECL generation conditions (e.g., voltage waveform, electrode type, amount of composition, amount of ECL label, etc.), produce substantially similar ECL signals as compositions containing the same components, except that a mild surfactant is present instead of the strict surfactant. In the embodiments, the strict surfactant is TRITON® X-100, TRITON® X-114, NP-40, IGEPAL® CA-630, 3-[(3-colamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), or sodium dodecyl sulfate (SDS).In the embodiments, the mild surfactant is a BRIJ®, TWEEN®, PLURONIC®, or KOLLIPHOR® surfactant, or an alkyl ether-PEG surfactant such as PEG(18) tridecyl ether.

[0062] In the embodiment, the concentration of alkyldiethanolamine or amphoteric tertiary amine in the composition is about 10 mM to about 500 mM, about 20 mM to about 400 mM, about 50 mM to about 250 mM, or about 100 mM to about 200 mM. In the embodiments, the concentration of the alkyldiethanolamine amphoteric tertiary amine in the composition is about 10 mM, about 20 mM, about 30 mM, about 40 mM, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 250 mM, about 300 mM, about 350 mM, about 400 mM, about 450 mM, or about 500 mM. In the embodiments, the alkyldiethanolamine is tBDEA. In the embodiments, the alkyldiethanolamine is MDEA. In the embodiments, the alkyldiethanolamine is a combination of tBDEA and MDEA. In the embodiment, the amphoteric tertiary amine is DEA-PS. In the embodiment, the composition comprises two or more combinations of tBDEA, MDEA, and DEA-PS.

[0063] In embodiments, a composition comprising an alkyldiethanolamine and / or an zwitterionic tertiary amine (e.g., tBDEA, MDEA, and / or DEA-PS) further comprises a pH buffering component. In embodiments, the pH buffering component has a pKa of from about 7.0 to about 8.0. In embodiments, the pH buffering component can maintain the pH of the composition at from about 7.0 to about 8.5, from about 7.2 to about 8.0, or from about 7.4 to about 7.9. In embodiments, the pH buffering component comprises Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, PIPES, MOPS, TES, DIPSO, MOBS, TAPSO, POPSO, HEPPSO, HEPPS, tricine, glycine amide, HE PBS, bicine, or a combination thereof. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the pH buffering component comprises Tris. In embodiments, the pH buffering component comprises phosphate.

[0064] In embodiments, the concentration of the pH buffering component in the composition is from about 10 mM to about 800 mM, from about 20 mM to about 600 mM, from about 50 mM to about 400 mM, from about 100 mM to about 300 mM, from about 120 mM to about 280 mM, or from about 150 mM to about 250 mM. In embodiments, the concentration of the pH buffering component in the composition is about 50 mM, about 100 mM, about 150 mM, about 200 mM, about 250 mM, about 300 mM, about 350 mM, about 400 mM, about 450 mM, or about 500 mM.

[0065] Ionic component In embodiments, the compositions herein comprise an ionic component. Ionic components such as salts dissociate into ions in solution. It has been found that high ionic concentrations can advantageously reduce non-specific binding of ECL labels to ECL co-reactants. Non-limiting examples of ionic components include cations Li + , Na + , K + , Rb + , Cs + , Mg +2 , Ca +2 , and NH4 +Salts containing, and / or anion F - Cl - , Br - , I - Examples include salts containing phosphates, sulfates, and borates. In embodiments, the ionic component is Li + na + , or K + It includes. In this embodiment, the ionic component is Cl - This includes. In embodiments, the ionic component includes lithium chloride (LiCl), sodium chloride (NaCl), potassium chloride (KCl), or a combination thereof. In embodiments, the ionic component includes NaCl. In embodiments, the ionic component includes KCl.

[0066] In the embodiment, the concentration of the ionic component in the composition is approximately 100 mM to approximately 2000 mM, approximately 200 mM to approximately 1800 mM, approximately 300 mM to approximately 1700 mM, approximately 400 mM to approximately 1600 mM, approximately 500 mM to approximately 1500 mM, approximately 600 mM to approximately 1200 mM, approximately 700 mM to approximately 1000 mM, or approximately 800 mM to approximately 900 mM. In the embodiment, the concentration of the ionic component in the composition is approximately 500 mM, approximately 550 mM, approximately 600 mM, approximately 650 mM, approximately 700 mM, approximately 750 mM, approximately 800 mM, approximately 850 mM, approximately 900 mM, approximately 950 mM, approximately 1000 mM, approximately 1100 mM, approximately 1200 mM, approximately 1300 mM, approximately 1400 mM, or approximately 1500 mM.

[0067] In the embodiment, the composition contains approximately 100 mM to approximately 2000 mM, approximately 200 mM to approximately 1800 mM, approximately 300 mM to approximately 1700 mM, approximately 400 mM to approximately 1600 mM, approximately 500 mM to approximately 1500 mM, approximately 600 mM to approximately 1200 mM, approximately 700 mM to approximately 1000 mM, or approximately 800 mM to approximately 900 mM of NaCl. In the embodiment, the composition contains KCl in concentrations of approximately 100 mM to 2000 mM, 200 mM to 1800 mM, 300 mM to 1700 mM, 400 mM to 1600 mM, 500 mM to 1500 mM, 600 mM to 1200 mM, 700 mM to 1000 mM, or 800 mM to 900 mM. In the embodiment, the composition contains approximately 100 mM to approximately 2000 mM, approximately 200 mM to approximately 1800 mM, approximately 300 mM to approximately 1700 mM, approximately 400 mM to approximately 1600 mM, approximately 500 mM to approximately 1500 mM, approximately 600 mM to approximately 1200 mM, approximately 700 mM to approximately 1000 mM, or approximately 800 mM to approximately 900 mM of LiCl.

[0068] In the embodiments, the composition has an ionic strength of about 0.2 M to about 2 M, about 0.5 M to about 1.5 M, about 0.75 M to about 1.25 M, or about 0.8 M to about 1.0 M. In the embodiments, the composition has an ionic strength of about 0.3 M or more, about 0.5 M or more, about 0.8 M or more, or about 1.0 M or more. In the embodiments, the composition contains chloride ions, and the concentration of chloride ions is about 0.3 M or more, about 0.5 M or more, about 0.8 M or more, or about 1.0 M or more.

[0069] In the embodiment, nonspecific binding (NSB) in an immunoassay using ECL as the assay readout is lower in a composition containing an ionic component compared to a composition that is identical except for not containing an ionic component.

[0070] surfactants When using compositions provided herein, for example, those containing TEA, tBDEA, MDEA, and / or DEA-PS as ECL co-reactants, it was unexpectedly discovered that the ECL-generating properties of the composition are substantially unaffected by the presence, concentration, or structure of the surfactant in the composition. In contrast, TPA-based compositions generally require the presence of a surfactant for optimal signal generation. In particular, TPA provides optimal ECL generation in the presence of a surfactant containing an aromatic moiety, such as the phenol ether moiety in TRITON® X-100.

[0071] In embodiments, the compositions herein are substantially free of surfactants. In embodiments, the compositions herein contain surfactants. In embodiments, the compositions herein contain surfactants at a concentration below the critical micelle concentration (CMC) of the surfactant. The CMC is the concentration of surfactant at which micelles are formed, and any additional amount of surfactant added to the composition above the CMC is incorporated into the micelles. The CMC of a surfactant can be determined by those skilled in the art, for example, using the titration method described in Wu et al., Anal Chem 92(6):4259-4265 (2020) and / or using a device such as a dynamic contact angle analyzer and / or a tensiometer.

[0072] In embodiments, the compositions herein include nonionic surfactants. In embodiments, the compositions herein include ionic surfactants. Examples of nonionic surfactants include NONIDET® (octylphenoxypolyethoxyethanol), BRIJ® (polyoxyethylene fatty acid ether), TRITON® (2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol), TWEEN® (polysorbate), KOLLIPHOR® (polyoxyl castor oil), THESIT® (polyethylene glycol dodecyl ether), LUBROL® (polyoxyethylene alkyl ether), and GENAPOL® (isotridecyl alcohol poly Examples of surfactants include glycol ethers, PLURONIC® (poloxamer block copolymer of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) arranged as PEO-PPO-PEO), TETRONIC® (poloxamine block copolymer of PEO-PPO), SYNPERONIC® (block copolymer of poly(ethylene glycol) (PEG) and poly(propylene glycol) (PPG) arranged as PEG-PPG-PEG), and SPAN® (sorbitan), which are known by trade names. Specific examples of nonionic surfactants include, for example, KOLLIPHOR® P-407 (PEG 101 -PPG 56 -PEG 101 (Also known as Poloxamer 407), PLURONIC (registered trademark) P-123 (PEO 18 -PPO 72 -PEO 18 ), PLURONIC(registered trademark) L-121 (PEG5-PPG 68 -PEG5), PLURONIC(registered trademark) 31R1(PPO 26 -PEO5-PPO 26Examples include alkyl ether-polyethylene glycol (PEG) such as ), TETRONIC® 701 (ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol), BRIJ® L4 (polyethylene glycol dodecyl ether), BRIJ® 58 (polyethylene glycol hexadecyl ether), TWEEN® 20 (polysorbate 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, and PEG(10) tridecyl ether, PEG(12) tridecyl ether, and PEG(18) tridecyl ether.

[0073] In embodiments, the surfactant comprises a phenol ether. In embodiments, the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100). In relation to the surfactants described herein, TRITON® X-100 is a “severe” surfactant capable of disrupting, dissolving, and / or dissolving lipid bilayer membranes, such as the membranes of cells or extracellular vesicles (EVs).

[0074] In the embodiments, the surfactant does not contain an aromatic moiety. In the embodiments, the surfactant does not contain a phenol ether. In the embodiments, the surfactant does not disrupt, lyse, or dissolve lipid bilayer membranes, such as cell or extracellular vesicle (EV) membranes. Such surfactants may be referred to as “mild” surfactants. Examples of mild surfactants include classes of surfactants known by trade names such as BRIJ®, TWEEN®, PLURONIC®, or KOLLIPHOR®. In the embodiments, the surfactant does not contain an ester linkage. In the embodiments, the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68-PEG5 (PLURONIC® L-121), PPO 26 -PEO5-PPO 26 (PLURONIC® 31R1), ethylene diamine tetrakis(propoxylate-block-ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20, 2,4,7,9-tetramethyl-d-decene-4,7-diol ethoxylate, alkyl ether-polyether-polyethylene glycol (PEG), or combinations thereof. In embodiments, the surfactant is an alkyl ether-polyethylene glycol (PEG). In embodiments, the alkyl ether-polyethylene glycol (PEG) is PEG(10) tridecyl ether, PEG(12) tridecyl ether, PEG(18) tridecyl ether, or combinations thereof. In embodiments, the surfactant is PEG(18) tridecyl ether.

[0075] As described herein, the compositions herein advantageously provide consistent ECL signal generation in the presence of different types of surfactants, such as the stringent surfactants and mild surfactants described herein. In embodiments, compositions comprising TEA, tBDEA, MDEA, DEA-PS, or combinations thereof and a stringent surfactant produce substantially similar ECL signals as compositions comprising the same components, except that a mild surfactant is present instead of the stringent surfactant, when subjected to the same ECL generation conditions (e.g., voltage waveform, type of electrode, amount of composition, amount of ECL label, etc.). In embodiments, the stringent surfactant is TRITON™ X-100. In embodiments, the mild surfactant is a BRIJ®, TWEEN®, PLURONIC®, or KOLLIPHOR® surfactant, or an alkyl ether-PEG surfactant such as PEG(18) tridecyl ether.

[0076] In an embodiment, the concentration of the surfactant in the composition is such that the composition has a gas-liquid surface tension of about 50 dynes / cm or less, about 40 dynes / cm or less, or about 35 dynes / cm or less. In an embodiment, the surfactant is present in the composition at its CMC, about 2 times or more its CMC, or about 5 times or more its CMC.

[0077] In an embodiment, the surfactant is about 0.1% (v), about 0.5% (v / v), about 1% (v / v), about 2% (v / v), about 5% (v / v), about 7% (v / v), or about 10% (v / v) of the composition. In an embodiment, the concentration of the surfactant in the composition is about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM. In an embodiment, the concentration of the surfactant in the composition is about 0.1 mM, about 0.2 mM, about 0.3 mM, about 0.4 mM, about 0.5 mM, about 0.6 mM, about 0.7 mM, about 0.8 mM, about 0.9 mM, about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM.

[0078] In an embodiment, the composition contains 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON™ X-100) at about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM. In an embodiment, the composition contains Poloxamer 407 (KOLLIPHOR® P-407) at about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM. In an embodiment, the composition contains PEO 18 -PPO 72 -PEO 18(PLURONIC® P-123) is included. In embodiments, the composition contains approximately 0.1 mM to 20 mM, approximately 0.1 mM to 10 mM, approximately 0.5 mM to 8 mM, approximately 0.75 mM to 6 mM, or approximately 1 mM to 5 mM of PEG5-PPG. 68 - Contains PEG5 (PLURONIC® L-121). In embodiments, the composition contains approximately 0.1 mM to approximately 20 mM, approximately 0.1 mM to approximately 10 mM, approximately 0.5 mM to approximately 8 mM, approximately 0.75 mM to approximately 6 mM, or approximately 1 mM to approximately 5 mM of PPO 26 -PEO5-PPO 26(PLURONIC® 31R1) is included. In embodiments, the composition contains about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM of ethylenediaminetetrakis(propoxylate-block-ethoxylate)tetrol (TETRONIC® 701). In embodiments, the composition contains about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM of polyethylene glycol dodecyl ether (BRIJ® L4). In the embodiments, the composition contains approximately 0.1 mM to approximately 20 mM, approximately 0.1 mM to approximately 10 mM, approximately 0.5 mM to approximately 8 mM, approximately 0.75 mM to approximately 6 mM, or approximately 1 mM to approximately 5 mM of polyethylene glycol hexadecyl ether (BRIJ® 58). In the embodiments, the composition contains approximately 0.1 mM to approximately 20 mM, approximately 0.1 mM to approximately 10 mM, approximately 0.5 mM to approximately 8 mM, approximately 0.75 mM to approximately 6 mM, or approximately 1 mM to approximately 5 mM of polysorbate 20 (TWEEN® 20). In the embodiments, the composition contains about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM of 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate. In the embodiments, the composition contains about 0.1 mM to about 20 mM, about 0.1 mM to about 10 mM, about 0.5 mM to about 8 mM, about 0.75 mM to about 6 mM, or about 1 mM to about 5 mM of alkyl ether-PEG. In the embodiments, the alkyl ether-PEG is PEG(18) tridecyl ether.

[0079] pH In embodiments, the compositions of this specification have a pH of about 6.0 to about 9.0, about 7.0 to about 8.0, about 7.2 to about 7.6, about 7.5 to about 7.8, about 7.4 to about 7.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, or about 8.0. In embodiments, the pH of the composition is about 7.5. In embodiments, the pH of the composition is about 7.8.

[0080] In the embodiment, the composition containing TEA has a pH of about 7.0 to about 8.0, about 7.4 to about 7.9, or about 7.5 to about 7.8 and is substantially free of additional pH buffering components. In the embodiment, the composition containing TEA has a pH of about 7.0 to about 8.0, about 7.4 to about 7.9, or about 7.5 to about 7.8 and is substantially free of additional components having a pKa of about 7.0 to about 8.0. In the embodiment, the composition containing TEA does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, and citrates. In the embodiment, the composition containing TEA does not contain any of HEPES, POPSO, HEPPSO, and PIPES. In the embodiment, the composition containing TEA does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES.

[0081] ECL labeled component In embodiments, the compositions of this specification include an ECL-labeled component. In embodiments, for example, an ECL co-reactant composition provided herein, comprising TEA, tBDEA, MDEA, and / or DEA-PS, and an ECL-labeled component, can generate ECL. In embodiments, the ECL-labeled component includes an ECL label. In embodiments, the ECL-labeled component includes a detection reagent. In embodiments, the ECL-labeled component includes a binding partner for the detection reagent.

[0082] In embodiments, the ECL-labeled component is a detection reagent containing an ECL label. In embodiments, the detection reagent comprises an antibody or its antigen detection fragment, an antigen, a ligand, a receptor, an oligonucleotide, a hapten, an epitope, a mimotope, or an aptamer. In embodiments, the detection reagent is an antibody or a variant thereof comprising its antigen / epitope detection portion, an antibody fragment or derivative, an antibody analog, a modified antibody, or a substance that binds to the antigen in a manner similar to that of an antibody. In embodiments, the detection reagent comprises at least one heavy-chain or light-chain complementarity-determining region (CDR) of an antibody. In embodiments, the detection reagent comprises at least two CDRs from one or more antibodies. In embodiments, the detection reagent is an antibody or its antigen detection fragment. In embodiments, the detection reagent is covalently linked to the ECL label via a conjugated linker. Methods for conjugating a label, such as an ECL label, to a detection reagent are known to those skilled in the art.

[0083] In the embodiment, the ECL-labeled component is a binding partner of the detection reagent. In the embodiment, the ECL-labeled component and the detection reagent form a complex that can be detected by ECL. In the embodiment, the ECL-labeled component and the detection reagent include receptor-ligand pairs, antigen-antibody pairs, hapten-antibody pairs, epitope-antibody pairs, mimotope-antibody pairs, aptamer-target molecule pairs, or intercalator-target molecule pairs. In the embodiment, the ECL-labeled component and the detection reagent include complementary oligonucleotides. In the embodiment, the ECL-labeled component and the detection reagent include biotin-avidin or biotin-streptavidin pairs.

[0084] In embodiments, the ECL label comprises an electrochemiluminescent organometallic complex. As used herein, the terms “electrochemiluminescence” and “ECL activity” may be used interchangeably. In embodiments, the electrochemiluminescent organometallic complex comprises ruthenium, osmium, iridium, rhenium, and / or lanthanide metals. In embodiments, the ECL label comprises ruthenium. In embodiments, the electrochemiluminescent organometallic complex comprises a substituted or unsubstituted bipyridine, or a substituted or unsubstituted phenanthroline. In embodiments, the ECL label comprises a substituted bipyridine. In embodiments, the ECL label comprises ruthenium(II) tris-bipyridine. In embodiments, the ECL label comprises an organometallic complex comprising at least one substituted bipyridine ligand, each substituted bipyridine ligand comprising at least one sulfonate group. In embodiments, the ECL label comprises an organometallic complex comprising at least two substituted bipyridine ligands, each substituted bipyridine ligand comprising at least one sulfonate group. In the embodiment, a substituted bipyridine ligand containing at least one sulfonate group is given by formula I: [ka] It is a compound of [the compound].

[0085] In one embodiment, the ECL label comprises three ligands, the first ligand being a compound of formula I, and the second ligand comprising a bipyridine having at least one substituent covalently attached to the detection reagent. In another embodiment, the ECL label comprises an organometallic complex comprising three ligands, two of which are compounds of formula I, and the third ligand comprising a bipyridine having at least one substituent covalently attached to the detection reagent.

[0086] In this embodiment, the first detectable marker is given by formula II: [ka] It is a compound of [the compound].

[0087] Additional exemplary ECL labels are found in US5,714,089, US6,136,268, US6,316,607, US6,468,741, US6,479,233, US6,808,939, and US9,499,573.

[0088] TEA composition In embodiments, the present invention provides a composition comprising from about 1000 mM to about 6500 mM TEA and from about 500 mM to about 1500 mM ionic component, wherein the composition has a pH of from about 7.0 to about 8.0 and the ionic component is NaCl, KCl, or LiCl. In embodiments, the composition substantially does not contain additional pH buffering components. In embodiments, the composition substantially does not contain additional components having a pKa of from about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL label component. In embodiments, the ECL label component is a detection reagent comprising an ECL label.

[0089] In embodiments, the present invention provides a composition comprising from about 1100 mM to about 3500 mM TEA and from about 600 mM to about 1200 mM ionic component, wherein the composition has a pH of from about 7.0 to about 8.0 and the ionic component is NaCl, KCl, or LiCl. In embodiments, the composition substantially does not contain additional pH buffering components. In embodiments, the composition substantially does not contain additional components having a pKa of from about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL label component. In embodiments, the ECL label component is a detection reagent comprising an ECL label.

[0090] In embodiments, the present invention provides a composition comprising about 1200 mM to about 1600 mM TEA and about 700 mM to about 900 mM of an ionic component, wherein the composition has a pH of about 7.0 to about 8.0 and the ionic component is NaCl, KCl, or LiCl. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label. In embodiments, the composition comprises about 1200 mM TEA and about 850 mM NaCl. In one embodiment, the composition contains about 1600 mM TEA and about 850 mM NaCl. In another embodiment, the composition contains about 1200 mM TEA and about 850 mM KCl. In another embodiment, the composition contains about 1600 mM TEA and about 850 mM KCl. In another embodiment, the composition contains about 1200 mM TEA and about 850 mM LiCl. In another embodiment, the composition contains about 1600 mM TEA and about 850 mM LiCl. In yet another embodiment, the composition has a pH of about 7.5. In yet another embodiment, the composition has a pH of about 7.8.

[0091] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM of TEA, about 500 mM to about 1500 mM of an ionic component, and about 0.1 mM to about 10 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0092] In embodiments, the present invention relates to a composition comprising about 1100 mM to about 3500 mM of TEA, about 600 mM to about 1200 mM of an ionic component, and about 0.5 mM to about 8 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26-PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0093] In embodiments, the present invention relates to a composition comprising about 1200 mM to about 1600 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.4 to about 7.9, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0094] In embodiments, the present invention relates to a composition comprising about 1200 mM TEA, about 850 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.4 to about 7.9, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26(PLURONIC® 31R1), ethylene diamine tetra (propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9 - tetramethyl - d - decyne - 4,7 - diol ethoxylate, alkyl ether - polyethylene glycol (PEG), or a combination thereof. In an embodiment, the surfactant is PEG(18) tridecyl ether. In an embodiment, the composition substantially does not contain additional pH buffering components. In an embodiment, the composition substantially does not contain additional components having a pKa of about 7.0 to about 8.0. In an embodiment, the composition does not contain any of Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, POPSO, HEPPSO, and PIPES. In an embodiment, the composition further contains an ECL labeling component. In an embodiment, the ECL labeling component is a detection reagent containing an ECL label.

[0095] In an embodiment, the present invention is a composition comprising about 1600 mM of TEA, about 850 mM of an ionic component, and about 1 mM of a surfactant, the composition having a pH of about 7.4 to about 7.9, the ionic component being NaCl, KCl, or LiCl, and the surfactant being 2 - [4 - (2,4,4 - trimethylpentan - 2 - yl)phenoxy]ethanol (TRITON™ X - 100), Poloxamer 407 (KOLLIPHOR® P - 407), PEO 18 -PPO 72 -PEO 18 (PLURONIC® P - 123), PEG5 - PPG 68 -PEG5 (PLURONIC® L - 121), PPO 26 -PEO5 - PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0096] In embodiments, the present invention relates to a composition comprising about 3200 mM TEA, about 850 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.4 to about 7.9, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0097] In embodiments, the present invention relates to a composition comprising about 6400 mM TEA, about 850 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.4 to about 7.9, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0098] In embodiments, the present invention relates to a composition comprising about 1200 mM TEA, about 700 mM to about 900 mM ionic components, and about 1 mM to about 5 mM surfactants, wherein the composition has a pH of about 7.0 to about 8.0, the ionic components are NaCl, KCl, or LiCl, and the surfactants are 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26(PLURONIC® 31R1), ethylene diamine tetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decyne-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In an embodiment, the surfactant is PEG(18) tridecyl ether. In an embodiment, the composition substantially does not contain additional pH buffering components. In an embodiment, the composition substantially does not contain additional components having a pKa of about 7.0 to about 8.0. In an embodiment, the composition does not contain any of Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, POPSO, HEPPSO, and PIPES. In an embodiment, the composition further comprises an ECL labeling component. In an embodiment, the ECL labeling component is a detection reagent containing an ECL label.

[0099] In an embodiment, the present invention provides a composition comprising about 1600 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON™ X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC® P-123), PEG5-PPG 68 -PEG5 (PLURONIC® L-121), PPO 26 -PEO5-PPO 26Provided is a composition that is (PLURONIC® 31R1), ethylene diamine tetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decyne-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In an embodiment, the surfactant is PEG(18) tridecyl ether. In an embodiment, the composition substantially does not contain an additional pH buffering component. In an embodiment, the composition substantially does not contain an additional component having a pKa of about 7.0 to about 8.0. In an embodiment, the composition does not contain any of Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, POPSO, HEPPSO, and PIPES. In an embodiment, the composition further comprises an ECL labeling component. In an embodiment, the ECL labeling component is a detection reagent comprising an ECL label.

[0100] In an embodiment, the present invention is a composition comprising about 3200 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON™ X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC® P-123), PEG5-PPG 68 -PEG5(PLURONIC® L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0101] In embodiments, the present invention relates to a composition comprising about 6400 mM TEA, about 700 mM to about 900 mM ionic components, and about 1 mM to about 5 mM surfactants, wherein the composition has a pH of about 7.0 to about 8.0, the ionic components are NaCl, KCl, or LiCl, and the surfactants are 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0102] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM TEA, about 850 mM NaCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0103] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM TEA, about 850 mM KCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0104] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM TEA, about 850 mM LiCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0105] In embodiments, the present invention provides a composition comprising about 1000 mM to about 6500 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100). In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0106] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0107] In embodiments, the present invention provides a composition comprising about 1000 mM to about 6500 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0108] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.5, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0109] In embodiments, the present invention relates to a composition comprising about 1000 mM to about 6500 mM of TEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.8, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In the embodiment, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0110] In one embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In an embodiment, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5.In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0111] In one embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In an embodiment, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5.In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0112] In one embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In an embodiment, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5.In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0113] In embodiments, the present invention relates to a composition comprising about 1200 mM TEA, about 850 mM NaCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM NaCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0114] In embodiments, the present invention relates to a composition comprising about 1600 mM TEA, about 850 mM NaCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM NaCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0115] In embodiments, the present invention relates to a composition comprising about 3200 mM TEA, about 850 mM NaCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM NaCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0116] In embodiments, the present invention relates to a composition comprising about 6400 mM TEA, about 850 mM NaCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM NaCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0117] In embodiments, the present invention relates to a composition comprising about 1200 mM TEA, about 850 mM KCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM KCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0118] In embodiments, the present invention relates to a composition comprising about 1600 mM TEA, about 850 mM KCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM KCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0119] In embodiments, the present invention relates to a composition comprising about 3200 mM TEA, about 850 mM KCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM KCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0120] In embodiments, the present invention relates to a composition comprising about 6400 mM TEA, about 850 mM KCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM KCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0121] In embodiments, the present invention relates to a composition comprising about 1200 mM TEA, about 850 mM LiCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM LiCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0122] In embodiments, the present invention relates to a composition comprising about 1600 mM TEA, about 850 mM LiCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM LiCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0123] In embodiments, the present invention relates to a composition comprising about 3200 mM TEA, about 850 mM LiCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM LiCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0124] In embodiments, the present invention relates to a composition comprising about 6400 mM TEA, about 850 mM LiCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM LiCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the compositions substantially contain no additional pH buffering components. In embodiments, the compositions substantially contain no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the compositions do not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the compositions further contain an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0125] In one embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In one embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In yet another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component.In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0126] In one embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In one embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In yet another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component.In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0127] In one embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 1200 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In one embodiment, the present invention provides a composition comprising about 1600 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In yet another embodiment, the present invention provides a composition comprising about 3200 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In embodiments, the present invention provides a composition comprising about 6400 mM TEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition substantially contains no additional components having a pKa of about 7.0 to about 8.0. In embodiments, the composition does not contain any of Tris, phosphates, HEPES, glycylglycine, borates, acetates, citrates, POPSO, HEPPSO, and PIPES. In embodiments, the composition further comprises an ECL-labeled component.In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0128] In embodiments, the present invention provides a composition comprising TEA, an ionic component, and optionally one or both of an ECL-labeled component and / or a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and optionally, the composition is substantially free of additional pH buffering components. In embodiments, the composition comprises about 1000 mM to about 6500 mM of TEA and about 500 mM to about 2000 mM of an ionic component. In embodiments, the surfactant comprises alkyl ether-PEG. In embodiments, the composition is substantially free of additional pH buffering components. In embodiments, the composition is essentially composed of or consists of the described components.

[0129] In embodiments, the present invention provides a composition comprising TEA, an ionic component, and one or both of an ECL-labeled component and a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and optionally, the composition is substantially free of additional pH buffering components. In embodiments, the composition comprises about 1000 mM to about 6500 mM of TEA and about 500 mM to about 2000 mM of an ionic component. In embodiments, the surfactant comprises alkyl ether-PEG. In embodiments, the composition is substantially free of additional pH buffering components. In embodiments, the composition is essentially composed of or consists of the described components.

[0130] In embodiments, the present invention provides a composition comprising TEA, an ionic component, and optionally one or both of an ECL-labeled component and / or a surfactant, wherein the composition has a pH of about 7.0 to about 8.0 and is substantially free of additional pH buffering components. In embodiments, the composition comprises about 1000 mM to about 6500 mM of TEA and about 500 mM to about 2000 mM of an ionic component. In embodiments, the surfactant comprises alkyl ether-PEG. In embodiments, the composition is essentially composed of or consists of the described components.

[0131] In embodiments, the present invention provides a composition comprising TEA, an ionic component, a surfactant, and an ECL-labeled component, wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the present invention provides a composition comprising TEA, an ionic component, and a surfactant, wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the present invention provides a composition comprising TEA, an ionic component, a surfactant, an ECL-labeled component, and a surfactant, wherein the composition has a pH of about 7.0 to about 8.0. In embodiments, the composition comprises about 1000 mM to about 6500 mM of TEA and about 500 mM to about 2000 mM of an ionic component. In embodiments, the surfactant comprises alkyl ether-PEG. In embodiments, the composition substantially contains no additional pH buffering components. In embodiments, the composition essentially consists of or comprises the described components.

[0132] In embodiments, the compositions provided herein are in a dry form. In embodiments, the compositions provided herein are in the form of a dry powder. In embodiments, the compositions provided herein are lyophilized powders. Throughout this disclosure, when a composition contains a specific concentration of its described components (e.g., about 1000 mM to about 6500 mM of TEA, about 500 mM to about 2000 mM of ionic components, and / or about 0.1 mM to about 10 mM of surfactants), and / or a specific pH of its described components (e.g., about 7.0 to about 8.0 pH), it will be understood by those skilled in the art that when a composition contains a specific concentration of its described components (e.g., about 1000 mM to about 6500 mM of TEA, about 500 mM to about 2000 mM of ionic components, and / or about 0.1 mM to about 10 mM of surfactants), and / or a specific pH of its described components (e.g., about 7.0 to about 8.0 pH), the described concentrations and pH of the components of the composition will be understood to relate to a composition in liquid form, such as a dry composition reconstituted with a liquid diluent (e.g., water or aqueous assay buffer). In embodiments, when a composition is in a dry form and reconstituted with a liquid diluent, it contains the described components at the described concentrations. In embodiments, when a composition is in a dry form and composed of a liquid diluent, it contains the described pH.

[0133] Alkyldiethanolamine / amphoterionic tertiary amine composition In embodiments, the present invention relates to a composition comprising about 50 mM to about 250 mM tBDEA, about 500 mM to about 1500 mM ionic component, and about 0.1 mM to about 10 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0134] In embodiments, the present invention relates to a composition comprising about 50 mM to about 250 mM of MDEA, about 500 mM to about 1500 mM of an ionic component, and about 0.1 mM to about 10 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0135] In embodiments, the present invention relates to a composition comprising about 50 mM to about 250 mM DEA-PS, about 500 mM to about 1500 mM ionic components, and about 0.1 mM to about 10 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic components are NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0136] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM tBDEA, about 700 mM to about 900 mM ionic component, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0137] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of MDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0138] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 700 mM to about 900 mM ionic components, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic components are NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0139] In embodiments, the present invention relates to a composition comprising about 150 mM tBDEA, about 850 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0140] In embodiments, the present invention relates to a composition comprising about 150 mM MDEA, about 850 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0141] In embodiments, the present invention relates to a composition comprising about 150 mM DEA-PS, about 850 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0142] In embodiments, the present invention relates to a composition comprising about 150 mM tBDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0143] In embodiments, the present invention relates to a composition comprising about 150 mM MDEA, about 700 mM to about 900 mM ionic component, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0144] In embodiments, the present invention relates to a composition comprising about 150 mM DEA-PS, about 700 mM to about 900 mM ionic component, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0145] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM tBDEA, about 850 mM NaCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0146] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM MDEA, about 850 mM NaCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0147] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 850 mM NaCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0148] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM tBDEA, about 850 mM KCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0149] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM MDEA, about 850 mM KCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0150] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 850 mM KCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0151] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM tBDEA, about 850 mM LiCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0152] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM MDEA, about 850 mM LiCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0153] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 850 mM LiCl, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0154] In an embodiment, the present invention provides a composition comprising about 100 mM to about 200 mM of tBDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100). In an embodiment, the present invention provides a composition comprising about 100 mM to about 200 mM of MDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100). In embodiments, the present invention provides a composition comprising about 100 mM to about 200 mM DEA-PS, about 700 mM to about 900 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100). In embodiments, the composition further comprises about 100 mM to about 200 mM pH buffering component. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0155] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of tBDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of MDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 700 mM to about 900 mM ionic components, and about 1 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic components are NaCl, KCl, or LiCl, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0156] In one embodiment, the present invention provides a composition comprising about 100 mM to about 200 mM of tBDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is PEG(18) tridecyl ether. In another embodiment, the present invention provides a composition comprising about 100 mM to about 200 mM of MDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM of a surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is PEG(18) tridecyl ether. In embodiments, the present invention provides a composition comprising about 100 mM to about 200 mM DEA-PS, about 700 mM to about 900 mM ionic component, and about 1 mM surfactant, wherein the composition has a pH of about 7.0 to about 8.0, the ionic component is NaCl, KCl, or LiCl, and the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises about 100 mM to about 200 mM pH buffering component. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0157] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of tBDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.5, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18(PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0158] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of tBDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.8, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0159] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of MDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.5, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0160] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM of MDEA, about 700 mM to about 900 mM of an ionic component, and about 1 mM to about 5 mM of a surfactant, wherein the composition has a pH of about 7.8, the ionic component is NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0161] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 700 mM to about 900 mM ionic components, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.5, the ionic components are NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0162] In embodiments, the present invention relates to a composition comprising about 100 mM to about 200 mM DEA-PS, about 700 mM to about 900 mM ionic components, and about 1 mM to about 5 mM surfactant, wherein the composition has a pH of about 7.8, the ionic components are NaCl, KCl, or LiCl, and the surfactant is 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the surfactant is PEG(18) tridecyl ether. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0163] In one embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM NaCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In yet another embodiment, the composition further comprises about 100 mM to about 200 mM of a pH buffering component. In the embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In the embodiments, the composition further comprises an ECL-labeled component.In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0164] In one embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM KCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the composition further comprises about 100 mM to about 200 mM of a pH buffering component. In the embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In the embodiments, the composition further comprises an ECL-labeled component.In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0165] In one embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In one embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.5. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM LiCl, and about 1 mM 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (TRITON® X-100), wherein the composition has a pH of about 7.8. In embodiments, the composition further comprises about 100 mM to about 200 mM of a pH buffering component. In the embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In the embodiments, the composition further comprises an ECL-labeled component.In the embodiment, the ECL-labeled component is a detection reagent containing an ECL label.

[0166] In embodiments, the present invention relates to a composition comprising about 150 mM tBDEA, about 850 mM NaCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM NaCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0167] In embodiments, the present invention relates to a composition comprising about 150 mM MDEA, about 850 mM NaCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM NaCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0168] In embodiments, the present invention relates to a composition comprising about 150 mM DEA-PS, about 850 mM NaCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM NaCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0169] In embodiments, the present invention relates to a composition comprising about 150 mM tBDEA, about 850 mM KCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM KCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0170] In embodiments, the present invention relates to a composition comprising about 150 mM MDEA, about 850 mM KCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM KCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0171] In embodiments, the present invention relates to a composition comprising about 150 mM DEA-PS, about 850 mM KCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM KCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0172] In embodiments, the present invention relates to a composition comprising about 150 mM tBDEA, about 850 mM LiCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM LiCl, and about 1 mM surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0173] In embodiments, the present invention relates to a composition comprising about 150 mM MDEA, about 850 mM LiCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM LiCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0174] In embodiments, the present invention relates to a composition comprising about 150 mM DEA-PS, about 850 mM LiCl, and about a surfactant, wherein the composition has a pH of about 7.5, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 The present invention provides compositions comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate)tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM LiCl, and about 1 part surfactant, wherein the composition has a pH of about 7.8, and the surfactant is Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26The present invention provides a composition comprising (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, alkyl ether-polyethylene glycol (PEG), or a combination thereof. In embodiments, the composition further comprises a pH buffering component of about 100 mM to about 200 mM. In embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In embodiments, the composition further comprises an ECL-labeled component. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0175] In one embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In one embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In yet another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM NaCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the composition further comprises about 100 mM to about 200 mM of a pH buffering component. In the embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In the embodiments, the composition further comprises an ECL-labeled component. In the embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0176] In one embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In one embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In yet another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM KCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the composition further comprises about 100 mM to about 200 mM of a pH buffering component. In the embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In the embodiments, the composition further comprises an ECL-labeled component. In the embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0177] In one embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In another embodiment, the present invention provides a composition comprising about 150 mM tBDEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In one embodiment, the present invention provides a composition comprising about 150 mM MDEA, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.5. In yet another embodiment, the present invention provides a composition comprising about 150 mM DEA-PS, about 850 mM LiCl, and about 1 mM PEG(18) tridecyl ether, wherein the composition has a pH of about 7.8. In yet another embodiment, the composition further comprises about 100 mM to about 200 mM of a pH buffering component. In the embodiments, the pH buffering component is Tris, phosphate, HEPES, glycylglycine, borate, acetate, citrate, or a combination thereof. In the embodiments, the composition further comprises an ECL-labeled component. In the embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0178] In one embodiment, the present invention provides a composition comprising an ECL co-reactant, an ionic component, and a surfactant. In the embodiment, the ECL co-reactant is tributylamine (TBA), (dibutyl)aminoethanol (DBAE), (diethyl)aminoethanol (DEAE), triethanolamine (TEA), butyldiethanolamine (BDEA), propyldiethanolamine (PDEA), ethyldiethanolamine (EDEA), methyldiethanolamine (MDEA), tert-butyldiethanolamine (tBDEA), dibutylamine (DBA), butylethanolamine (BEA), diethanolamine (DEA), dibutylamine propyl sulfonate (DBA-PS), dibutylamine butyl sulfonate (DBA-BS), butylethanolamine propyl sulfonate (BEA-PS), butylethanolamine butyl sulfonate (BEA-BS), diethanolamine propyl sulfonate (also known as 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid, DEA-PS), diethanolamine butyl sulfonate (DEA-BS), or a combination thereof. In the embodiments, the composition has a pH of about 7.0 to about 8.0. In the embodiments, the composition further comprises a pH buffering component. Suitable ionic components (e.g., NaCl, KCl, and LiCl), surfactants (e.g., TRITON X-100 or mild surfactants as described herein), pH buffering components (e.g., Tris or phosphates), and their concentrations in the composition are provided herein. In the embodiments, the composition further comprises an ECL-labeled component. In the embodiments, the ECL-labeled component is a detection reagent containing an ECL label.

[0179] method In embodiments, the present invention provides a method for generating electrochemiluminescence (ECL), comprising (a) contacting an electrode with an ECL co-reactant composition provided herein, (b) applying a voltage to the electrode, and (c) generating ECL. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the present invention provides a method for generating electrochemiluminescence (ECL), comprising (a) contacting an electrode with a TEA composition comprising TEA, an ionic component, and optionally a surfactant, (b) applying a voltage to the electrode, and (c) generating ECL. In embodiments, the method further comprises detecting the generated ECL. In embodiments, the method further comprises measuring the generated ECL. In embodiments, the electrode is located on a surface.

[0180] In embodiments, the present invention provides a method for generating electrochemiluminescence (ECL), comprising (a) contacting an electrode with (i) an ECL co-reactant composition provided herein and (ii) an ECL label, (b) applying a voltage to the electrode, and (c) generating ECL. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the present invention provides a method for generating electrochemiluminescence (ECL), comprising (a) contacting an electrode with (i) a TEA composition comprising TEA, an ionic component, and optionally a surfactant, and (ii) an ECL label, (b) applying a voltage to the electrode, and (c) generating ECL. In embodiments, the method further comprises detecting the generated ECL. In embodiments, the method further comprises measuring the generated ECL to quantify the amount of the ECL label. In embodiments, the electrode is located on a surface.

[0181] In embodiments, the present invention provides a method for quantifying the amount of ECL label in a sample, comprising: (a) contacting an electrode with (i) an ECL co-reactant composition or a TEA composition provided herein, and (ii) a sample containing the ECL label; (b) applying a voltage to the electrode; (c) generating ECL; (d) measuring the ECL; and (e) quantifying the amount of ECL label from the measured ECL. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the TEA composition includes TEA, an ionic component, and optionally a surfactant.

[0182] In the embodiments, ECL is produced from a reaction between an ECL co-reactant (e.g., TEA, tBDEA, MDEA, and / or DEA-PS) in the composition herein and an ECL label. In the embodiments, the ECL label is present on the ECL label component. In the embodiments, the ECL label is present in the sample. In the embodiments, the sample contains the ECL label component. In the embodiments, the ECL label component contains a detection reagent. In the embodiments, the sample contains the binding partner of the ECL label component. In the embodiments, the ECL label component contains the binding partner of the detection reagent. Detection reagents and their binding partners are further described herein. In the embodiments, the ECL label component is present in a binding complex, and the method further comprises detecting the binding complex by detecting the ECL produced. In the embodiments, the method comprises contacting an electrode with a sample containing the binding partner of the ECL label component, the ECL label component and the binding partner form a binding complex, and the method further comprises detecting the binding complex by detecting the ECL produced. In the embodiment, the method includes measuring the generated ECL and thereby quantifying the amount of the ECL-labeled component and / or the bound complex.

[0183] In the embodiments, each of the sample, the ECL co-reacting material composition or TEA composition provided herein, and the ECL-labeled component is dry. In the embodiments, each of the sample, the ECL co-reacting material composition or TEA composition provided herein, and the ECL-labeled component is liquid. In the embodiments, one or more of the sample, the ECL co-reacting material composition or TEA composition provided herein, and the ECL-labeled component are dry, and the remaining components are liquid. For example, the sample is liquid, and one or both of the ECL co-reacting material composition or TEA composition and the ECL-labeled component provided herein are dry. In embodiments including a liquid component and a dry component, the liquid component reconstitutes the dry component. In embodiments, the method further includes contacting the electrode with a liquid diluent to reconstitute the dry component in the liquid. In embodiments, the dry component is present on the surface. In embodiments, the ECL co-reacting material composition is dry and present on the surface. In embodiments, the TEA composition is dry and present on the surface. In embodiments, the ECL-labeled component is dry and present on the surface. Compositions in dry form are described herein. In embodiments, the ECL-labeled component is a detection reagent containing an ECL label. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the TEA composition includes TEA, an ionic component, and optionally a surfactant.

[0184] In embodiments, the ECL-labeled component in the binding complex is a first copy of the detection reagent containing the ECL label. In embodiments, the binding complex comprises a first copy of the detection reagent and a binding reagent immobilized on a surface. The binding reagent is further described herein. In embodiments, the method further comprises forming a binding complex. In embodiments, the binding complex is formed before or during step (a) of the method.

[0185] In the embodiment, the conjugated complex is formed by incubating an assay mixture comprising a conjugating reagent, a first copy of the detection reagent, and a second copy of the detection reagent containing an ECL label, under conditions that the conjugated complex is formed on a surface and the second copy of the detection reagent remains in solution. In the embodiment, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In the embodiment, the ECL co-reactant composition comprises TEA. In the embodiment, the TEA composition comprises TEA, an ionic component, and optionally a surfactant.

[0186] In the embodiments, the conjugated complex is formed by incubating an assay mixture comprising a conjugating reagent, a first copy of the detection reagent, a second copy of the detection reagent including an ECL label, and an ECL co-reactant composition or TEA composition provided herein, under conditions such that the conjugated complex is formed on a surface and the second copy of the detection reagent remains in solution. In the embodiments, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In the embodiments, the composition comprises TEA. In the embodiments, the TEA composition comprises TEA, an ionic component, and optionally a surfactant.

[0187] In the embodiments, the conjugated complex is formed by combining the sample with a first copy of the detection reagent, a second copy of the detection reagent including an ECL label, and an ECL co-reactant composition or TEA composition provided herein to form an assay mixture, and by contacting the assay mixture with the conjugated reagent under conditions that the conjugated complex is formed on the surface and the second detection reagent remains in solution. In the embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In the embodiments, the composition includes TEA. In the embodiments, the TEA composition includes TEA, an ionic component, and optionally a surfactant.

[0188] In the embodiments, the binding complex further comprises an analyte, which is further described herein. In the embodiments, the binding reagent and the detection reagent each specifically bind to the analyte. In the embodiments, the method includes detecting the analyte by detecting the generated ECL. In the embodiments, the method includes measuring the generated ECL to quantify the amount of the analyte.

[0189] In the embodiment, the ECL label comprises an electrochemiluminescent organometallic complex. In the embodiment, the electrochemiluminescent organometallic complex comprises ruthenium, osmium, iridium, rhenium, and / or lanthanide metals. In the embodiment, the ECL label comprises ruthenium. In the embodiment, the ECL label comprises ruthenium(II) tris-bipyridine. In the embodiment, the electrochemiluminescent organometallic complex comprises a substituted or unsubstituted bipyridine, or a substituted or unsubstituted phenanthroline. In the embodiment, the ECL label comprises a substituted bipyridine. In the embodiment, the ECL label comprises an organometallic complex comprising at least one substituted bipyridine ligand, each substituted bipyridine ligand comprising at least one sulfonate group. In the embodiment, the ECL label comprises an organometallic complex comprising at least two substituted bipyridine ligands, each substituted bipyridine ligand comprising at least one sulfonate group. In the embodiment, the substituted bipyridine ligand comprising at least one sulfonate group is a compound of formula I. In the embodiment, the ECL label comprises the compound of formula II.

[0190] In embodiments, the compositions herein are used, for example, in an ECL-based binding assay to detect and / or quantify an analyte of interest and / or a binding complex containing the analyte. In embodiments, the binding complex is formed, for example, on a surface including an electrode, and the binding complex includes an ECL label that can generate ECL when in contact with an ECL co-reactant described herein. Binding assays include, but are not limited to, the following: (1) a direct binding assay in which the analyte of interest is labeled with an ECL label, a binding reagent which is the binding partner of the analyte is immobilized on the surface, and a binding complex is formed by direct binding of the binding reagent and the labeled analyte; (2) a sandwich binding assay in which both the immobilized binding reagent and the detection reagent containing the ECL label are binding partners of the analyte, and the analyte binds to the two binding partners to form a binding complex; (3) a competitive binding assay in which the immobilized binding reagent is the binding partner of the analyte and the labeled detection reagent is a competitor (e.g., the analyte or a structural analog of the analyte) that competes with the immobilized binding reagent for binding to the analyte, or the labeled detection reagent is the binding partner of the analyte and the immobilized binding reagent is a competitor that competes with the detection reagent for binding to the analyte. In a competitive binding assay, the amount of the labeled binding complex formed by direct binding of the immobilized binding reagent and the labeled detection reagent decreases as the amount of analyte increases. Binding assays are further described, for example, in WO2014 / 165061, WO2014 / 160192, WO2015 / 175856, US9,618,510, US10,114,015, US10,408,823, US2017 / 0168047, and US2019 / 0011441.

[0191] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) contacting a liquid sample with a surface containing an ECL co-reactant or TEA composition provided herein, wherein the liquid sample contains an ECL-labeled component or a binding partner of an ECL-labeled component, and the method further comprises contacting the surface with the ECL-labeled component, thereby forming a bound complex containing the ECL-labeled component on the surface; (b) applying a voltage to the surface to generate ECL; and (c) detecting the generated ECL, thereby detecting the bound complex. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the ECL co-reactant composition includes TEA. In embodiments, the TEA composition includes TEA, an ionic component, and optionally a surfactant. In embodiments, the surface includes an electrode. In embodiments, the ECL-labeled component includes a detection reagent containing an ECL label. In the embodiments, the ECL-labeled component comprises a detection reagent containing an ECL label, and the binding complex comprises the binding reagent and the detection reagent. In the embodiments, the ECL-labeled component comprises a binding partner of the detection reagent, and the binding partner contains an ECL label. In the embodiments, the ECL-labeled component comprises a binding partner of the detection reagent, and the binding complex comprises the binding reagent, the detection reagent, and the binding partner. The detection reagent and the binding partner are further described herein. In the embodiments, the detection reagent and the ECL-labeled component comprise complementary oligonucleotides. In the embodiments, the binding complex further comprises an analyte. In the embodiments, the binding reagent and the detection reagent each bind specifically to the analyte.

[0192] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) contacting a liquid sample with a surface, the surface comprising an ECL-labeled component and an ECL co-reactant composition or TEA composition provided herein, the liquid sample comprising a binding partner of the ECL-labeled component, thereby forming a bound complex containing the ECL-labeled component on the surface; (b) applying a voltage to the surface to generate ECL; and (c) detecting the generated ECL, thereby detecting the bound complex. In embodiments, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the ECL co-reactant composition comprises TEA. In embodiments, the TEA composition comprises TEA, an ionic component, and optionally a surfactant. In embodiments, the surface comprises an electrode. In embodiments, the ECL-labeled component comprises a detection reagent containing an ECL label. In embodiments, the ECL-labeled component comprises a detection reagent containing an ECL label, and the bound complex comprises the binding reagent and the detection reagent. In the embodiments, the ECL-labeled component includes a binding partner of the detection reagent, and the binding partner includes an ECL label. In the embodiments, the ECL-labeled component includes a binding partner of the detection reagent, and the binding complex includes a binding reagent, a detection reagent, and a binding partner. The detection reagent and the binding partner are further described herein. In the embodiments, the detection reagent and the ECL-labeled component include a complementary oligonucleotide. In the embodiments, the binding complex further includes an analyte. In the embodiments, the binding reagent and the detection reagent each specifically bind to the analyte.

[0193] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) forming a bound complex on a surface, wherein the bound complex comprises an ECL-labeled component; (b) contacting the bound complex with an ECL co-reactant composition or TEA composition provided herein; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex. In embodiments, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the ECL co-reactant composition comprises TEA. In embodiments, the TEA composition comprises TEA, an ionic component, and optionally a surfactant. In embodiments, the surface comprises an electrode. In embodiments, the ECL-labeled component comprises a detection reagent containing an ECL label. In embodiments, the ECL-labeled component comprises a detection reagent containing an ECL label, and the bound complex comprises the binding reagent and the detection reagent. In the embodiments, the ECL-labeled component includes a binding partner of the detection reagent, and the binding partner includes an ECL label. In the embodiments, the ECL-labeled component includes a binding partner of the detection reagent, and the binding complex includes a binding reagent, a detection reagent, and a binding partner. The detection reagent and the binding partner are further described herein. In the embodiments, the detection reagent and the ECL-labeled component include a complementary oligonucleotide. In the embodiments, the binding complex further includes an analyte. In the embodiments, the binding reagent and the detection reagent each specifically bind to the analyte.

[0194] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) forming a bound complex on a surface, wherein the surface includes an electrode, and the bound complex comprises a binding reagent immobilized on the surface and a detection reagent including an electrochemiluminescence (ECL) label; (b) contacting the bound complex with an ECL co-reactant composition or TEA composition provided herein; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex. In embodiments, the bound complex further comprises an analyte, and the binding reagent and detection reagent each specifically bind to the analyte. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the ECL co-reactant composition includes TEA. In embodiments, the TEA composition includes TEA, an ionic component, and optionally a surfactant. In embodiments, the bound complex further comprises an analyte. In this embodiment, the binding reagent and the detection reagent each bind specifically to the analyte.

[0195] In embodiments, the present invention provides a method for detecting a target analyte in a sample, comprising: (a) contacting the sample with a surface containing a binding reagent, the binding reagent being specifically bound to the analyte; and (ii) a detection reagent being specifically bound to the analyte, the detection reagent comprising an electrochemiluminescence (ECL) label, thereby forming a binding complex on the surface comprising the binding reagent, the analyte, and the detection reagent; (b) contacting the binding complex on the surface with an ECL co-reactant composition or TEA composition provided herein; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL. In embodiments, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In embodiments, the ECL co-reactant composition includes TEA. In embodiments, the TEA composition includes TEA, an ionic component, and optionally a surfactant. In embodiments, the surface includes an electrode.

[0196] As discussed herein, the compositions provided herein may be used in ECL-based assays that do not require a washing step. In the context of surface-based ECL assays, the “washing step” refers to adding a washing buffer to a surface to remove undesirable components from the assay reaction mixture, such as excess, nonspecifically bound, or unbound reagents (e.g., detection reagents and / or ECL labels) from the sample, and / or unbound or nonspecifically bound components from the sample. In one example, a biological sample may contain the analyte of interest and various other biomaterials that are not of interest and do not specifically bind to the binding reagent, and the washing step can remove such components from the reaction mixture. In embodiments, the composition comprises TEA. In embodiments, the composition comprises TEA, an ionic component, and optionally a surfactant. In a “washing” assay, the washing step is typically used to remove unbound ECL labels before detecting ECL labels on a surface. The washing step may be eliminated if the detection method can effectively distinguish between surface-bound ECL labels (e.g., as part of the bound complex being detected) or unbound "free" ECL labels in solution. "No-wash" assay forms, which eliminate the washing step, are often advantageous because performing the washing step can be difficult or cumbersome in many situations. However, no-wash assay forms are typically difficult to develop due to high background ECL signals resulting from incomplete differentiation between free and bound ECL labels present in the reaction mixture. Even in assays that utilize a washing step, good differentiation between bound and free ECL labels is advantageous to provide greater robustness against washing inefficiencies or variations in washing quality by offering tolerance to low levels of free label contamination that may be associated with poor washing.

[0197] As discussed herein, the compositions herein have remarkably distinguished between free ECL labels and bound ECL labels in ECL-based assays performed on solid surfaces (e.g., solid electrode surfaces). In embodiments, the compositions herein increase the ratio of ECL signals from bound labels to ECL signals from free labels. Thus, the compositions herein provide improved assay performance, particularly when measuring low affinity interactions, which requires the presence of high concentrations of ECL labels in the reaction, but which are also expected to suffer significant signal loss due to the dissociation of bound complexes during the washing step. In embodiments, the compositions comprise TEA. In embodiments, the compositions comprise TEA, an ionic component, and optionally a surfactant.

[0198] Although not constrained by theory, the compositions and ECL co-reactants (e.g., TEA) described herein are thought to reduce the distance from the solid electrode surface at which ECL is generated from the ECL label. This increases the signal from the bound label (held close to the electrode) compared to the free label (distributed throughout the solution on the electrode). The increase in signal from the bound label can also be characterized in terms of "effective excitation length," which is the maximum distance at which the free ECL label can be excited. The "effective excitation length" is influenced by: (1) the distance of short intermediates involved in the generation of ECL (e.g., oxidation products of the ECL co-reactant) that can diffuse from the electrode before being depleted by side reactions (a function of the lifetime and diffusion constant of these intermediates); and (2) the rate at which the free label or unbound reagent diffuses into a region close enough to the electrode to participate in the reaction with these reactive intermediates (a function of the diffusion constant of the unbound ECL label or labeled reagent). In methods using the compositions of this specification, the effective excitation length is reduced by more than 2, 3, 4, 5, 9, 7, 8, 9, or 10 times compared to compositions containing TPA. In embodiments, the composition contains TEA. In embodiments, the composition contains TEA, an ionic component, and optionally a surfactant.

[0199] In embodiments, the methods herein do not include a washing step. In embodiments where the method detects a bound complex, the method does not include a washing step before, during, or after the formation of the bound complex on a surface. In embodiments where the method detects an analyte of interest in a sample, the method does not include a washing step before, during, or after contacting the sample with (i) a surface containing a binding reagent on which the binding reagent specifically binds to the analyte, and (ii) a detection reagent that specifically binds to the analyte. In embodiments, the method does not include a washing step before, during, or after contacting the bound complex with the composition. In embodiments, the method does not include a washing step before, during, or after applying a voltage to the surface to generate ECL. In embodiments, the method does not include a washing step before or during detection of the generated ECL. In embodiments, the composition includes TEA. In embodiments, the composition includes TEA, an ionic component, and optionally a surfactant.

[0200] In embodiments, the methods herein include a washing step. In embodiments where the method detects a bound complex, the method includes a washing step before, during, or after the formation of a bound complex on a surface. In embodiments where the method detects an analyte of interest in a sample, the method includes a washing step before, during, or after contacting the sample with (i) a surface containing a bounding reagent on which the bounding reagent specifically binds to the analyte, and (ii) a detection reagent that specifically binds to the analyte. In embodiments, the method includes a washing step before, during, or after contacting the bound complex with the composition. In embodiments, the method includes a washing step before, during, or after applying a voltage to the surface to generate ECL. In embodiments, the method includes a washing step before or during detection of the generated ECL. In embodiments, the composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof.

[0201] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) forming an assay mixture by combining a sample with a detection mixture comprising (i) an ECL co-reactant composition or TEA composition provided herein and (ii) at least two copies of a detection reagent, each copy of the detection reagent comprising an ECL label; (b) contacting the assay mixture with a bound reagent immobilized on a surface including an electrode, under conditions that (I) a bound complex is formed on the surface, the bound complex comprises the bound reagent and a first copy of the detection reagent, and (II) a second copy of the detection reagent remains in solution; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex. In embodiments, the surface includes an electrode. In embodiments, the second copy of the detection reagent is not removed before any of steps (b) to (d). In embodiments, the second copy of the detection reagent is not removed before step (b). In the embodiment, the ECL co-reactant composition includes TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In the embodiment, the ECL co-reactant composition includes TEA. In the embodiment, the TEA composition includes TEA, an ionic component, and optionally a surfactant.

[0202] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) an assay mixture comprising (i) a bound reagent immobilized on a surface, the surface optionally comprising an electrode; and (ii) a detection mixture comprising at least two copies of a detection reagent, each copy of the detection reagent comprising an electrochemiluminescence (ECL) label; (i) incubation under conditions that a bound complex is formed on the surface, the bound complex comprising the bound reagent and a first copy of the detection reagent, and (ii) a second copy of the detection reagent remaining in solution; (b) contacting the bound complex with an ECL co-reactant composition or TEA composition provided herein; (c) applying a voltage to the surface to generate ECL; and (d) detecting the generated ECL to detect the bound complex. In embodiments, the surface comprises an electrode. In embodiments, the method further comprises washing the surface before any of steps (b) to (d) to remove the second copy of the detection reagent. In the embodiment, the second copy of the detection reagent is not removed before any of steps (b) to (d). In the embodiment, the second copy of the detection reagent is not removed before step (b). In the embodiment, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In the embodiment, the ECL co-reactant composition comprises TEA. In the embodiment, the TEA composition comprises TEA, an ionic component, and optionally a surfactant.

[0203] In embodiments, the present invention provides a method for detecting a bound complex, comprising: (a) an assay mixture comprising (i) a bound reagent immobilized on a surface, the surface optionally comprising an electrode; (ii) a detection mixture comprising at least two copies of a detection reagent, each copy of the detection reagent comprising an electrochemiluminescent (ECL) label; and (iii) an ECL co-reactant composition or TEA composition provided herein; (i) a bound complex formed on the surface, the bound complex comprising the bound reagent and a first copy of the detection reagent, and (ii) incubation under conditions that a second copy of the detection reagent remains in solution; (b) a voltage applied to the surface to generate ECL; and (c) detection of the generated ECL to detect the bound complex. In embodiments, the surface comprises an electrode. In embodiments, the method further comprises washing the surface before either step (b) or (c) to remove the second copy of the detection reagent. In the embodiment, the second copy of the detection reagent is not removed before either step (b) or (d). In the embodiment, the second copy of the detection reagent is not removed before step (b). In the embodiment, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof. In the embodiment, the ECL co-reactant composition comprises TEA. In the embodiment, the TEA composition comprises TEA, an ionic component, and optionally a surfactant.

[0204] In the embodiment, the binding complex further comprises the analyte, and the first copies of the binding reagent and the detection reagent each specifically bind to the analyte.

[0205] In embodiments, at least two copies of the conjugating reagent are immobilized on the surface, and the first copy of the conjugating reagent binds to the competitor such that it complexes with the first copy of the detection reagent, while the second copy of the conjugating reagent binds to the competitor such that it does not complex with the second copy of the detection reagent. In embodiments, at least two copies of the conjugating reagent are immobilized on the surface, and the first copy of the conjugating reagent binds to the competitor such that it complexes with the first copy of the detection reagent, while the second copy of the detection reagent binds to the competitor such that the second copy of the conjugating reagent does not complex with the second copy of the detection reagent. Competitors and competitive assay formats are further described herein.

[0206] In the embodiment, the binding reagent binds to a first copy of the detection reagent to form a binding complex.

[0207] In embodiments, the conjugating reagent comprises an antibody or its antigen-binding fragment, an antigen, a ligand, a receptor, an oligonucleotide, a hapten, an epitope, a mimotope, or an aptamer. In embodiments, the conjugating reagent is an antibody or a variant thereof comprising its antigen / epitope-binding moiety, an antibody fragment or derivative, an antibody analog, a modified antibody, or a substance that binds to the antigen in a manner similar to that of an antibody. In embodiments, the conjugating reagent comprises at least one heavy-chain or light-chain complementarity-determining region (CDR) of an antibody. In embodiments, the conjugating reagent comprises at least two CDRs from one or more antibodies. In embodiments, the conjugating reagent is an antibody or its antigen-binding fragment. In embodiments, the conjugating reagent binds specifically to the analyte. As used herein, “specifically binds” means that the reagent (e.g., the conjugating reagent) preferentially binds to its binding partner (e.g., the epitope of the analyte) compared to random unrelated substances. In embodiments, the conjugating reagent is an antibody or its antigen-binding fragment comprising a binding domain that specifically binds to the epitope of the analyte.

[0208] In embodiments, the binding reagent is immobilized on the surface. In embodiments, the binding reagent is directly immobilized on the surface. In embodiments, the binding reagent is indirectly immobilized on the surface via the binding reagent and a secondary binding partner on the surface. Exemplary secondary binding partners include, but are not limited to, complementary oligonucleotides, receptor-ligand pairs, antigen-antibody pairs, hapten-antibody pairs, epitope-antibody pairs, mimotope-antibody pairs, aptamer-target molecule pairs, hybridization partners, intercalator-target molecule pairs, and interreactive moieties (e.g., thiols and maleimides or iodoacetamides, aldehydes and hydrazides, or azides and alkynes or cycloalkynes).

[0209] In embodiments, the detection reagent comprises an antibody or its antigen detection fragment, an antigen, a ligand, a receptor, an oligonucleotide, a hapten, an epitope, a mimotope, or an aptamer. In embodiments, the detection reagent is an antibody or a variant thereof comprising its antigen / epitope detection portion, an antibody fragment or derivative, an antibody analog, a modified antibody, or a substance that binds to the antigen in a similar manner to an antibody. In embodiments, the detection reagent comprises at least one heavy chain or light chain complementarity determining region (CDR) of an antibody. In embodiments, the detection reagent comprises at least two CDRs from one or more antibodies. In embodiments, the detection reagent is an antibody or its antigen detection fragment. In embodiments, the detection reagent specifically binds to the analyte. In embodiments, the detection reagent is an antibody or its antigen-binding fragment comprising a binding domain that specifically binds to an epitope in the analyte. In embodiments, the detection reagent binds to an epitope of an analyte different from that of the binding reagent. In embodiments, both the binding reagent and the detection reagent are antibodies or their antigen-binding fragments.

[0210] In the embodiment, the detection reagent includes an ECL label. In the embodiment, the ECL label includes an electrochemiluminescent organometallic complex. In the embodiment, the organometallic complex includes ruthenium, osmium, iridium, rhenium, and / or lanthanide metals. In the embodiment, the organometallic complex includes a substituted or unsubstituted bipyridine, or a substituted or unsubstituted phenanthroline. In the embodiment, the ECL label includes ruthenium. In the embodiment, the ECL label includes ruthenium(II) tris-bipyridine. In the embodiment, the ECL label includes a substituted bipyridine. In the embodiment, the ECL label includes an organometallic complex containing at least one substituted bipyridine ligand, each substituted bipyridine ligand containing at least one sulfonate group. In the embodiment, the ECL label includes an organometallic complex containing at least two substituted bipyridine ligands, each substituted bipyridine ligand containing at least one sulfonate group. In the embodiments, the substituted bipyridine ligand containing at least one sulfonate group is a compound of formula I. In the embodiments, the ECL label comprises a compound of formula II. Exemplary ECL labels are provided in US5,714,089, US6,136,268, US6,316,607, US6,468,741, US6,479,233, US6,808,939, and US9,499,573.

[0211] In embodiments, the binding reagent and / or detection reagent bind directly to the analyte. For example, the binding reagent and / or detection reagent are antibodies or antigen-binding fragments thereof that specifically bind to epitopes on the analyte, respectively. In embodiments, the binding reagent and / or detection reagent bind indirectly to the analyte via secondary interactions. In embodiments, the analyte is linked to the binding partner of the binding reagent and / or detection reagent. For example, the binding reagent and / or detection reagent contains streptavidin, and the analyte is linked to biotin. Further examples of binding partners that can be recognized by secondary interactions include, for example, avidin-biotin, streptavidin-biotin, antibody-hapten, antibody-epitope tag, nucleic acid-complementary nucleic acid, aptamer-aptamer target, and receptor-ligand.

[0212] In the embodiment, the surface includes a multiwell plate. In the embodiment, the surface includes particles. In the embodiment, the surface includes an assay cartridge. In the embodiment, the surface includes the surface of a slide, tip, well, assay cell or flow cell, tube, channel, beads, or microparticles. In the embodiment, the surface includes particles, and the method further includes collecting the particles onto an additional surface and applying a voltage to the particles on the additional surface. In the embodiment, the particles are beads (such as magnetic beads), and the method further includes collecting the beads onto a magnetized plate, wherein the plate includes electrodes, and applying a voltage to the plate. In the embodiment, the surface and / or additional surface includes electrodes. In the embodiment, the electrodes are carbon electrodes, platinum electrodes, gold electrodes, or silver electrodes. In the embodiment, the electrodes are carbon ink electrodes.

[0213] In embodiments, the method includes measuring the amount of the target analyte or binding complex in a sample. An approach using a measured ECL signal to determine the amount and / or concentration of the ECL label (or analyte or binding complex) in an ECL-based binding assay is known to those skilled in the art and includes, for example, establishing a relationship between the ECL signal and the amount and / or concentration of the ECL label and / or analyte using a calibration standard and / or calibration curve. Calibration may be performed at different times, for example, during method development, during qualification of a particular lot of assay material, and / or at the time of assay measurement. Calibration may also be performed using calculations based on the known physical and chemical behavior of the assay components and measuring instruments.

[0214] The methods described herein can be used to test various samples that may contain the analyte of interest. In embodiments, the sample is a biological sample. In the embodiments, the sample includes (live or dead) cells, immortalized cells, cell-derived products, cell fragments, cell fractions, (fractionated or unfractionated) cell lysates, eukaryotic cells, prokaryotic cells, organelles, cell nuclei and their fractions, cell membranes, hybridomas, cell culture supernatants (e.g., supernatants from antibody-producing organisms such as hybridomas), cytoskeleton, protein complexes, structural biological components, skeletal components such as ligaments and tendons, hair, fur, feathers, hair fractions, skin, dermis, endothelium, mammalian fluids, secretions, excrement, whole blood, plasma, serum, sputum, tears, lymph, synovial fluid, pleural fluid, urine, sweat, cerebrospinal fluid, ascites, milk, feces, bronchial lavage, saliva, amniotic fluid, nasal discharge, vaginal secretions, swab biopsies, sperm, semen / seminal The sample is derived from a fluid, wound secretions and excretions, mucosal swabs, tissue aspirates, tissue homogenates, or extractions, purifications, or dilutions thereof. In embodiments, the sample is derived from plants, plant by-products, soil, water sources, oils, wastewater, or environmental samples. In embodiments, the sample further comprises water, organic solvents (e.g., acetonitrile, dimethyl sulfoxide, dimethylformamide, n-methylpyrrolidone, alcohols, or combinations thereof), EDTA, heparin, citrates, or combinations thereof. The sample may be obtained from a single source as described herein or may contain mixtures from two or more sources.

[0215] The analytes that can be measured using the method of the present invention include, but are not limited to, whole cells, cell surface antigens, intracellular particles (e.g., organelles or membrane fragments), exosomes, extracellular vesicles, liposomes, membrane vesicles, viruses, prions, dust mites or their fragments, viroids, antibodies, antigens, haptens, fatty acids, nucleic acids (and synthetic analogs), proteins (and synthetic analogs), lipoproteins, polysaccharides, inhibitors, cofactors, haptens, cell receptors, receptor ligands, lipopolysaccharides, glycoproteins, peptides, polypeptides, enzymes, enzyme substrates, enzyme products, second messengers, cell metabolites, hormones, pharmacological agents, synthetic organic molecules, organometallic molecules, tranquilizers, barbiturates, alkaloids, steroids, vitamins, amino acids, sugars, lectins, recombinant proteins or derived proteins, biotin, avidin, streptavidin, or inorganic molecules present in the sample. Measurable activities include, but are not limited to, those of phosphorylases, phosphatases, esterases, trans-glutaminases, nucleic acid damage activity, transferases, oxidases, reductases, dehydrogenases, glycosidases, ribosomes, protein-processing enzymes (e.g., proteases, kinases, protein phosphatases, ubiquitin-protein ligases, etc.), nucleic acid-processing enzymes (e.g., polymerases, nucleases, integrases, ligases, helicases, telomerases, etc.), cell receptor activation, and activation of the second messenger system.

[0216] All cells can be animals, plants, or bacteria, and can be living or dead cells. Examples include fungi and plant pathogens such as nematodes. The term “intracellular particles” means to include, for example, intracellular organelles, membrane particles from destroyed cells, cell wall fragments, ribosomes, multi-enzyme complexes, and other particles that may originate from living organisms. Examples of nucleic acids include chromosomal DNA, plasmid DNA, viral DNA, and recombinant DNA from multiple sources. Examples of nucleic acids include RNA, such as messenger RNA, ribosomal RNA, and transfer RNA. Examples of polypeptides include structural proteins such as enzymes, transport proteins, receptor proteins, and viral coat proteins. In embodiments, polypeptides are enzymes or antibodies. In embodiments, polypeptides are monoclonal antibodies. Examples of hormones include insulin and T4 thyroid hormone. Pharmacological agents include, for example, cardiac glycosides. It is within the scope of this disclosure to include synthetic polypeptides, synthetic nucleic acids, and synthetic materials that are chemically similar to biological materials such as synthetic membranes, vesicles, and liposomes. The foregoing is not intended to be a comprehensive list of biological substances suitable for use in this disclosure, but rather to illustrate the broad scope of this disclosure.

[0217] In embodiments, the method described herein is a multiplexing method capable of detecting multiple binding complexes and / or analytes. In embodiments, the multiplexing method detects multiple binding complexes and / or analytes simultaneously. In embodiments, the multiplexing method comprises repeating one or more method steps to measure multiple binding complexes and / or analytes. In embodiments, each method step is performed in parallel for each binding complex and / or analyte. In embodiments where the method detects multiple binding complexes, each binding complex comprises a different binding reagent and / or detection reagent. In embodiments where the method detects multiple analytes, each analyte binds to a different binding reagent and / or detection reagent. In embodiments, the binding of each analyte to its corresponding binding reagent is performed in parallel by bringing the surface of a sample containing the multiple analytes into contact with the sample.

[0218] In the embodiments, the multiplexing method does not include a washing step. Multiplexed non-washing assays are particularly difficult because the amount of detection reagent present in the assay mixture increases, and therefore the amount of ECL label in the solution increases, contributing to a high background ECL signal. The ECL co-reactant described herein has remarkably good discrimination between bound and free ECL labels in multiplexed assay forms, including multiplexed non-washing assays, providing a high ECL signal and low background. In the embodiments, the ECL co-reactant is TEA.

[0219] In the embodiment, the surface includes multiple binding domains, and each binding complex is formed in a different binding domain. In the embodiment, the multiple binding domains are on a single surface. In the embodiment, the surface includes a multiwell plate, and each binding domain is in a different well. In the embodiment, the surface includes wells of a multiwell plate, and each binding domain is in a distinct portion of a well. In the embodiment, the multiple binding domains are on one or more surfaces. In the embodiment, the surface includes particles, and each binding domain is on a different particle. In the embodiment, the particles are arranged in a particle array. In the embodiment, the particles are coded to allow identification of specific particles and to distinguish each binding domain.

[0220] In the embodiment, each binding domain contains a targeting agent that can bind to a targeting agent complement, and each binding reagent contains an auxiliary binder that can bind to a binder. In the embodiment, the binding reagent is immobilized within the binding domain by (1) binding the binding reagent to a targeting agent complement connected to the binder via the auxiliary binder, and (2) binding the product of (1) to a binding domain containing a targeting agent, wherein (i) each binding domain contains a different targeting agent, and (ii) each targeting agent complement selectively binds to one of the targeting reagents.

[0221] In the embodiment, an optional crosslinking agent, which is a binding partner for both the linking agent and the auxiliary linking agent, crosslinks the linking agent and the auxiliary linking agent such that the binding reagents, each bound to their respective targeting agent complements, come into contact with the binding domains and bind to their respective targeting agents via the crosslinking agent, the targeting agent complements on each binding reagent, and the targeting agents on each binding domain.

[0222] In the embodiment, the targeting agent and targeting agent complement are two members of a binding partner pair selected from avidin-biotin, streptavidin-biotin, antibody-hapten, antibody-antigen, antibody-epitope tag, nucleic acid-complementary nucleic acid, aptamer-aptamer target, and receptor-ligand. In the embodiment, the targeting agent and targeting agent complement are cross-reactive moieties, for example, thiol and maleimide or iodoacetamide, aldehyde and hydrazide, or azide and alkyne or cycloalkyne. In the embodiment, the targeting agent is biotin, and the targeting agent complement is avidin or streptavidin.

[0223] In embodiments, the linker and auxiliary linker are two members of a binding partner pair selected from avidin-biotin, streptavidin-biotin, antibody-hapten, antibody-antigen, antibody-epitope tag, nucleic acid-complementary nucleic acid, aptamer-aptamer target, and receptor-ligand. In embodiments, the linker and auxiliary linker are cross-reactive moieties, such as thiol and maleimide or iodoacetamide, aldehyde and hydrazide, or azide and alkyne or cycloalkyne. In embodiments, the linker is avidin or streptavidin, and the auxiliary linker is biotin. In embodiments, the targeting agent and targeting agent complement are complementary oligonucleotides. In embodiments, the targeting agent complement is streptavidin, the targeting agent is biotin, and the linker and auxiliary linker are complementary oligonucleotides.

[0224] In embodiments containing a crosslinking agent, the crosslinking agent is streptavidin or avidin, and the binder and auxiliary binder are biotin, respectively.

[0225] In embodiments, the present invention provides a method for producing a composition comprising combining an ECL co-reactant, an ionic component, and a surfactant. In the embodiment, the ECL co-reactant is tributylamine (TBA), (dibutyl)aminoethanol (DBAE), (diethyl)aminoethanol (DEAE), triethanolamine (TEA), butyldiethanolamine (BDEA), propyldiethanolamine (PDEA), ethyldiethanolamine (EDEA), methyldiethanolamine (MDEA), tert-butyldiethanolamine (tBDEA), dibutylamine (DBA), butylethanolamine (BEA), diethanolamine (DEA), dibutylamine propyl sulfonate (DBA-PS), dibutylamine butylsulfonate (DBA-BS), butylethanolamine propylsulfonate (BEA-PS), butylethanolamine butylsulfonate (BEA-BS), diethanolamine propylsulfonate (DEA-PS), or diethanolamine butylsulfonic acid (DEA-BS, also known as 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid).

[0226] In embodiments, the present invention further provides a method for producing a composition comprising combining triethanolamine (TEA) and an ionic component. In embodiments, the present invention further provides a method for producing a composition comprising combining triethanolamine (TEA), an ionic component, and a surfactant, wherein the method does not involve the addition of an additional pH buffering component. In embodiments, one or more of the components are provided in a dry form. Suitable ionic components and surfactants for the composition are provided herein, for example, NaCl, KCl, and LiCl (in the case of an ionic component), as well as Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68-PEG5 (PLURONIC (registered trademark) L-121), PPO 26 -PEO5-PPO 26 This includes (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, and alkyl ether-polyethylene glycol (e.g., PEG(18) tridecyl ether) (in the case of surfactants). TEA, ionic components, and surfactants may be included in the concentrations specified herein. In embodiments, the composition produced by this method includes about 1000 mM to about 6500 mM of TEA, about 700 to about 1000 mM of ionic components, and about 0.5 mM to about 10 mM of surfactants. In this embodiment, the composition produced by this method contains about 1200 mM to about 1600 mM of TEA, about 700 to about 1000 mM of ionic components, and about 1 mM to about 5 mM of surfactant.

[0227] In embodiments, the present invention further provides a method for producing a composition comprising combining tert-butyldiethanolamine (tBDEA), methyldiethanolamine (MDEA), 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid (DEA-PS), or a combination thereof, an ionic component, and a surfactant. In embodiments, one or more of the components are provided in a dry form. Suitable ionic components and surfactants for the composition are provided herein, for example, NaCl, KCl, and LiCl (in the case of an ionic component), as well as Poloxamer 407 (KOLLIPHOR® P-407), PEO 18 -PPO 72 -PEO 18 (PLURONIC(registered trademark) P-123), PEG5-PPG 68 -PEG5 (PLURONIC (registered trademark) L-121), PPO26 -PEO5-PPO 26 This includes (PLURONIC® 31R1), ethylenediaminetetrakis(propoxylate block ethoxylate) tetrol (TETRONIC® 701), polyethylene glycol dodecyl ether (BRIJ® L4), polyethylene glycol hexadecyl ether (BRIJ® 58), polysorbate 20 (TWEEN® 20), 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, and alkyl ether-polyethylene glycol (e.g., PEG(18) tridecyl ether) (in the case of surfactants). tBDEA and / or MDEA, ionic components, and surfactants may be included in the concentrations specified herein. In this embodiment, the composition produced by this method contains about 50 mM to about 250 mM tBDEA, about 50 mM to about 250 mM MDEA and / or about 50 mM to about 250 mM DEA-PS, about 700 to about 1000 mM ionic components, and about 0.5 mM to about 10 mM surfactant.

[0228] Assay module In embodiments, the present invention provides an assay module comprising a TEA composition in a dry form, wherein the TEA composition comprises TEA, an ionic component, and optionally a surfactant. In embodiments, the present invention provides an assay module comprising an ECL co-reactant composition provided herein in a dry form. In embodiments, the ECL co-reactant composition comprises TEA, tBDEA, MDEA, DEA-PS, or a combination thereof.

[0229] In the embodiment, the assay module includes a multiwell plate. In the embodiment, the assay module includes an assay cartridge. In the embodiment, the assay module includes slides, tips, wells, assay cells or flow cells, tubes, channels, beads, or microparticles. In the embodiment, the assay module includes electrodes. In the embodiment, the electrodes are carbon electrodes, platinum electrodes, gold electrodes, or silver electrodes. In the embodiment, the electrodes are carbon ink electrodes.

[0230] In the embodiments, the assay module further comprises a conjugating reagent in dry form. In the embodiments, the assay module further comprises a detection reagent in dry form. In the embodiments, the assay module further comprises a conjugating reagent and a detection reagent in dry form. The conjugating reagent and the detection reagent are further described herein. In the embodiments, the detection reagent comprises an ECL label.

[0231] In the embodiment, the ECL label comprises an electrochemiluminescent organometallic complex. In the embodiment, the organometallic complex comprises ruthenium, osmium, iridium, rhenium, and / or lanthanide metals. In the embodiment, the organometallic complex comprises a substituted or unsubstituted bipyridine, or a substituted or unsubstituted phenanthroline. In the embodiment, the ECL label comprises ruthenium. In the embodiment, the ECL label comprises ruthenium(II) tris-bipyridine. In the embodiment, the ECL label comprises a substituted bipyridine. In the embodiment, the ECL label comprises an organometallic complex comprising at least one substituted bipyridine ligand, each substituted bipyridine ligand comprising at least one sulfonate group. In the embodiment, the ECL label comprises an organometallic complex comprising at least two substituted bipyridine ligands, each substituted bipyridine ligand comprising at least one sulfonate group. In the embodiment, the substituted bipyridine ligand comprising at least one sulfonate group is a compound of formula I. In the embodiment, the ECL label comprises the compound of formula II.

[0232] kit In embodiments, the present invention includes a kit comprising an ECL co-reactant composition or TEA composition as described herein. In the embodiment, the ECL co-reactant composition includes an ECL co-reactant selected from tributylamine (TBA), (dibutyl)aminoethanol (DBAE), (diethyl)aminoethanol (DEAE), triethanolamine (TEA), butyldiethanolamine (BDEA), propyldiethanolamine (PDEA), ethyldiethanolamine (EDEA), methyldiethanolamine (MDEA), tert-butyldiethanolamine (tBDEA), dibutylamine (DBA), butylethanolamine (BEA), diethanolamine (DEA), dibutylamine propyl sulfonate (DBA-PS), dibutylamine butyl sulfonate (DBA-BS), butylethanolamine propyl sulfonate (BEA-PS), butylethanolamine butyl sulfonate (BEA-BS), diethanolamine propyl sulfonate (DEA-PS, also known as 3-[bis-(2-hydroxy-ethyl)-amino]-propane-1-sulfonic acid), diethanolamine butyl sulfonate (DEA-BS), and combinations thereof. In the embodiment, the composition contains TEA. In the embodiment, the composition contains tBDEA. In the embodiment, the composition contains MDEA. In the embodiment, the composition contains MDEA. In the embodiment, the composition contains DEA-PS. In the embodiment, the TEA composition contains TEA, an ionic component, and optionally a surfactant.

[0233] In embodiments, the present invention provides a kit comprising two or more components that, when mixed, form a composition described herein. In embodiments, the present invention provides a kit comprising (a) triethanolamine (TEA) and (b) an ionic component in one or more containers, vials, or compartments. In embodiments, the present invention provides a kit comprising (a) triethanolamine (TEA) and (b) an ionic component in one or more containers, vials, or compartments, wherein the kit does not contain additional pH buffering components. In embodiments, the present invention provides a kit comprising (a) triethanolamine (TEA), (b) an ionic component, and (c) a surfactant in one or more containers, vials, or compartments, wherein the kit does not contain additional pH buffering components. Ionic components (e.g., NaCl, KCl, and / or LiCl), surfactants (e.g., TRITON X-100, KOLLIPHOR® P-407, PLURONIC® P-123, PLURONIC® L-121, PLURONIC® 31R), TETRONIC® 701, BRIJ® L4, BRIJ® 58, TWEEN® 20, 2,4,7,9-tetramethyl-d-decine-4,7-diol ethoxylate, and / or alkyl ether-polyethylene glycol (e.g., PEG(18) tridecyl ether)), and / or their concentrations are described herein.

[0234] In embodiments, the kit further comprises an assay reagent, a calibration reagent, a surface, an ECL label, or a combination thereof. In embodiments, the kit comprises an assay reagent. In embodiments, the assay reagent comprises a conjugate reagent, a detection reagent, or both. The conjugate reagent and the detection reagent are further described herein and include, for example, an antibody or its antigen-binding fragment, an antigen, a ligand, a receptor, an oligonucleotide, a hapten, an epitope, a mimotope, or an aptamer. In embodiments, the conjugate reagent is an antibody or its antigen-binding fragment. In embodiments, the detection reagent is an antibody or its antigen-binding fragment. In embodiments, both the conjugate reagent and the detection reagent are antibodies or their antigen-binding fragments.

[0235] In embodiments, the kit includes an assay module as described herein. In embodiments, the assay module includes an ECL co-reactant composition or TEA composition as described herein in a dry form. In embodiments, the kit includes a surface. Surfaces suitable for performing ECL-based binding assays are described herein. In embodiments, the surface includes a multi-well plate. In embodiments, the surface includes an assay cartridge. In embodiments, the surface includes particles. In embodiments, the surface includes a slide, tip, well, assay cell or flow cell, tube, bead, or microparticle surface. In embodiments where the surface includes particles, beads, or microparticles, the kit further includes an additional surface, such as a plate, for collecting the particles, beads, or microparticles. In embodiments, the additional surface includes magnetically collectible particles, beads, or microparticles. In embodiments, the additional surface further includes a magnetic plate. In embodiments, the surface and / or additional surface includes an electrode. In embodiments, the electrode is a carbon electrode, platinum electrode, gold electrode, or silver electrode. In embodiments, the electrode is a carbon ink electrode.

[0236] In embodiments, the binding reagent is immobilized on a surface. In embodiments, the binding reagent and the surface are provided separately in a kit, and the kit further includes reagents for immobilizing the binding reagent on the surface. Methods for immobilizing the binding reagent on a surface are provided herein and include, for example, direct or indirect immobilization via the binding reagent and a secondary binding partner on the surface.

[0237] In embodiments, the kit includes an ECL label. The ECL label is further described herein and includes, for example, a ruthenium-containing compound. In embodiments, the ECL label includes an electrochemiluminescent organometallic complex. In embodiments, the organometallic complex includes ruthenium, osmium, iridium, rhenium, and / or lanthanide metals. In embodiments, the organometallic complex includes a substituted or unsubstituted bipyridine, or a substituted or unsubstituted phenanthroline. In embodiments, the ECL label includes ruthenium. In embodiments, the ECL label includes ruthenium(II) tris-bipyridine. In embodiments, the ECL label includes a substituted bipyridine. In embodiments, the ECL label includes an organometallic complex comprising at least one substituted bipyridine ligand, each substituted bipyridine ligand comprising at least one sulfonate group. In embodiments, the ECL label includes an organometallic complex comprising at least two substituted bipyridine ligands, each substituted bipyridine ligand comprising at least one sulfonate group. In the embodiments, the substituted bipyridine ligand containing at least one sulfonate group is a compound of formula I. In the embodiments, the ECL label contains a compound of formula II. In the embodiments, the detection reagent contains the ECL label. In the embodiments, the detection reagent and the ECL label are provided separately in a kit, the kit further comprising a reagent for conjugating the detection reagent with the ECL label. Conjugation methods are known to those skilled in the art.

[0238] In embodiments, the kit includes a calibration reagent. In embodiments, the calibration reagent includes a known amount of the analyte of interest. In embodiments, the calibration reagent includes a known amount of the ECL label. In embodiments, the kit includes multiple calibration reagents containing the analyte or ECL label in a range of concentrations. In embodiments, the multiple calibration reagents include the analyte or ECL label in concentrations close to the upper and lower limits of quantification of the ECL-based binding assay described herein. In embodiments, the multiple calibration reagents cover the entire dynamic range of the binding assay. In embodiments, the calibration reagent is a positive control reagent. In embodiments, the calibration reagent is a negative control reagent. In embodiments, the positive or negative control reagent is used to provide a basis for comparison between the method of the present invention and the sample being tested.

[0239] In the embodiment, one or more components of the kit are provided in a dry form, for example, as a lyophilized reagent. In the embodiment, one or more components of the kit are provided in a solution. In the embodiment, the conjugating reagent is lyophilized. In the embodiment, the conjugating reagent is provided in a solution. In the embodiment, the detection reagent is lyophilized. In the embodiment, the detection reagent is provided in a solution. In the embodiment, the calibration reagent is lyophilized. In the embodiment, the calibration reagent is provided in a solution. In the embodiment, the kit further comprises a liquid diluent. In the embodiment, the liquid diluent reconstitutes the dry reagent. In the embodiment, the liquid diluent is water. In the embodiment, one or more components of the kit are provided as a concentrated stock solution, for example, at 2, 4, 5, 10, or 20 times the working concentration of the reagent.

[0240] In embodiments, the kit includes, for example, an assay instrument for detecting ECL produced from the compositions and methods described herein. In embodiments, the kit further includes assay consumables, such as an assay module configured to contain a sample and / or reagent during one or more steps of the method described herein; pipette tips and other consumables for transferring liquid samples and reagents; covers and seals for the assay module and other consumables used in the assay (e.g., tubes, cuvettes, wells, multiwell plates, cartridges, lateral flow devices, flow cells); racks for holding other assay consumables; labels for identifying samples (including human-readable or machine-readable formats such as barcodes and RFID); or other assay consumables and media (including paper and electronic media) for providing information about the method and / or instructions for performing the method.

[0241] All references cited herein, including patents, patent applications, papers, textbooks, etc., and references cited in them to the extent that they do not yet exist, are incorporated herein by reference in their entirety. [Examples]

[0242] Example 1. Evaluation of amphoteric ions and hydroxyethylamine ECL co-reactants The following ECL co-reactants were tested for their ability to distinguish surface-bound and free (solution-based) ECL labels in ECL generation and solid-surface ECL assays: tributylamine (TBA), (dibutyl)aminoethanol (DBAE), (diethyl)aminoethanol (DEAE), triethanolamine (TEA), butyldiethanolamine (BDEA), propyldiethanolamine (PDEA), ethyldiethanolamine (EDEA), methyldiethanolamine (MDEA), tert-butyldiethanolamine (tBDEA), dibutylamine (DBA), butylethanolamine (BEA), diethanolamine (DEA), dibutylamine propyl sulfonate (DBA-PS), dibutylamine butyl sulfonate (DBA-BS), butylethanolamine propyl sulfonate (BEA-PS), butylethanolamine butyl sulfonate (BEA-BS), diethanolamine propyl sulfonate (DEA-PS), and diethanolanolamine butyl sulfonate (DEA-BS). Each ECL reading buffer composition was prepared with 150 mM of a specified ECL co-reactant, 200 mM of phosphate, 850 mM of NaCl, and either TRITON® X-100 ("TX100") or PEG(18) tridecyl ether ("PEG18 TDE"), and adjusted to pH 7.5.

[0243] 2nM IgG conjugated with biotin and ECL labeling ("BTI") was used as a bound labeling control and brought into contact with a streptavidin-coated electrode surface. 500 mM free ECL labeling ("FT") was used as a free labeling control. The results are shown in Figures 1A and 1B. Figure 1A shows the ECL signals measured with BTI, FT, and background signal ("D100") using only ECL reading buffer (without labeling). Figure 1B shows the ratio of the ECL signal from bound labeling to the ECL signal from free labeling ("BTI / FT") and the signal-to-background ratio ("S / B").

[0244] The raw values ​​and ratios in Figures 1A and 1B indicate information regarding radical lifetime, excited state formation efficiency in both oxidation and reduction pathways, and reduction / oxidation quench efficiency of ECL-labeled excited states. The ECL signal sensitivity in TRITON® X-100 is related to short-lived amine radical cations or the -1 oxidation state (labeled). -1 ) supports low electron transfer efficiency to ECL labeling.

[0245] From the results, we can conclude that DBA-BS is sensitive to TRITON(trademark) X-100 and produces significantly more signal from the free label than from the bound label, suggesting that a long-lived reducing radical was produced, leading to efficient reduction of the free ECL label. Furthermore, the BTI / FT signal ratio of MDEA was higher than that of PIPES, an ECL co-reactant known to have a short radical cation lifetime, and MDEA had a reasonable signal-to-background ratio, but only in the presence of TRITON(trademark) X-100, suggesting that MDEA also has a short radical cation lifetime. BDEA showed a strong signal from BTI and an intermediate amount of signal from FT, which suggests a radical cation and shortens the radical lifetime between that of TBA and TEA. Notably, TEA showed a very low FT signal and a considerably high BTI signal, was insensitive to the presence or absence of TRITON(trademark) X-100, and had the highest BTI / FT ratio among all ECL co-reactants tested.

[0246] Example 2. Ratio of bound / free labeled signal to TEA concentration Because TEA has a pKa of 7.7, it has the ability to function as both a pH buffer and an ECL co-reactant. Various TEA concentrations from 50 mM to 1600 mM were tested for their ECL formation properties. Each ECL reading buffer composition tested contained a specific concentration of TEA and 850 mM NaCl (pH 7.8). The compositions were tested using BTI and FT labeling in the same manner as in Example 1.

[0247] The results are shown in Figures 2A-2C. Figure 2A shows plots of ECL and BTI / FT ratio generated from BTI and FT at different TEA concentrations. The dashed lines at the top and bottom of the plot represent the BTI signal and BTI / FT ratio generated using PIPES ECL reading buffer, respectively. Thus, TEA showed a peak BTI / FT ratio at a concentration of approximately 1200 mM, and at TEA concentrations above 1200 mM, the BTI signal was within 10-25% of that of PIPES ECL reading buffer. Decreased TEA radical cations and radical lifetimes, as well as changes in buffer viscosity, may have contributed to the general BTI / FT behavior and the apparent decrease at 1600 mM TEA. Figure 2B shows the measured ECL signals from BTI, FT, and background (D100) at different TEA concentrations, and Figure 2C shows the BTI / FT ratio, S / B ratio, and ECL generation percentage compared to PIPES ECL reading buffer.

[0248] Example 3. ECL signal versus co-reactant concentration The results of Example 2 suggested that increasing the concentration of TEA produced a higher ECL signal, which contradicted predictions based on the known behavior of other ECL co-reactants such as PIPES. The ECL signals produced using PIPES ECL reading buffer and TEA ECL reading buffer were measured with various concentrations of co-reactants. PIPES compositions contained 20 mM, 40 mM, or 80 mM PIPES, >0.1% TRITON® X-100, and 80–320 mM potassium phosphate buffer (pH 7.5). TEA compositions contained 50 mM, 100 mM, or 200 mM TEA, 850 mM NaCl, and 1 mM PEG18 TDE. The compositions were tested with BTI as described for the previous examples.

[0249] The results are shown in Figure 3A (ECL signal vs. PIPES concentration) and Figure 3B (relative ECL signal vs. PIPES concentration and TEA concentration plotted together). Figure 3A confirms that for the PIPES ECL read buffer, the ECL signal decreases as the PIPES concentration increases. Figure 3B shows the unexpected contrasting behavior of TEA, which, despite its short radical lifetime, shows a strong increase in the ECL signal with increasing TEA concentration.

[0250] Example 4. Different assay formats Figures 4A–4D show four different assay formats tested using ECL read buffers containing different ECL co-reactants: TPA, BDEA, PIPES, and 1.2M TEA. The assays were evaluated on a panel of analytes. Figures 4E–4H illustrate multiplexed versions of the assays in Figures 4A–4D.

[0251] Figure 4A shows a "standard" two-step washing assay in which a capture antibody ("cAb", binding reagent) immobilized on a binding domain ("BD") on a surface is brought into contact with a mixture of analytes (one of which specifically binds to the capture antibody), and the surface is then washed, resulting in the capture of the analytes on the surface. Next, a mixture of detection antibodies ("dAb", detection reagent), each containing an ECL label and one of which specifically binds to the analyte, is added to the surface, and the surface is then washed, resulting in a binding complex containing cAb, the analyte, and dAb. Next, ECL reading buffer is added to the surface, and the resulting ECL is read by an ECL reading instrument. Figure 4E shows a multiplexed version of the "standard" two-step washing assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody that can bind to an analyte in the analyte mixture. Washing the surface containing the binding domains after adding the analyte mixture results in the capture of multiple analytes on the binding domains. Next, a mixture of detection antibodies, each containing an ECL label and capable of binding to the analyte in the analyte mixture, is added to the surface, and then the surface is washed, resulting in several binding complexes, each binding complex containing cAb, the analyte, and dAb. Then, ECL reading buffer is added to the surface, and the resulting ECL is read by an ECL instrument.

[0252] In the examples herein using a standard two-step assay format, 50 μL of the analyte mixture was added to a plate and shaken at 705 rpm for 2 hours at room temperature. The plate was washed once with wash buffer, 25 μL of the detection antibody mixture was added to the plate, and shaken at 705 rpm for 1.5 hours at room temperature. The plate was washed once with wash buffer, and 150 μL of ECL reading buffer was added to the plate. The plate was then read using an ECL reader.

[0253] Figure 4B shows a "one-step" assay in which a capture antibody on a binding domain on a surface is brought into contact with the analyte mixture, and then the surface is washed as shown in Figure 4A. Next, the detection antibody mixture is added, followed by the addition of ECL reading buffer, but no washing is performed between the addition of the detection antibody mixture and the ECL reading buffer. The resulting ECL is then read by an ECL reader. Figure 4F shows a multiplexed version of the "one-step" assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody capable of binding to an analyte in the analyte mixture. The surfaces containing the binding domains are washed after adding the analyte mixture as shown in Figure 4E. The detection antibody mixture is added to form multiple binding complexes, and then the ECL reading buffer is added, but no washing is performed between the addition of the detection antibody mixture and the ECL reading buffer. The resulting ECL is then read by an ECL reader.

[0254] In the examples herein using a one-step assay format, 50 μL of the analyte mixture was added to a plate and shaken at 705 rpm for 2 hours at room temperature. The plate was washed once with washing buffer, 25 μL of the detection antibody mixture was added to the plate, and shaken at 705 rpm for 1.5 hours at room temperature. 125 μL of ECL reading buffer was added to the plate. The plate was then read using an ECL reader.

[0255] Figure 4C shows a "one-step no-wash" assay in which a capture antibody on a binding domain on a surface is brought into contact with the analyte mixture and the detection antibody mixture, and then into contact with ECL reading buffer, without washing between any steps. The resulting ECL is then read by an ECL reader. Figure 4G shows a multiplexed version of the "one-step no-wash" assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody capable of binding to an analyte in the analyte mixture. The surfaces containing the binding domains are brought into contact with the analyte mixture and the detection antibody mixture to form multiple binding complexes, and then ECL reading buffer is added, without washing between any steps. The resulting ECL is then read by an ECL reader.

[0256] In the examples herein using a one-step, no-wash assay format, 25 μL of the analyte mixture was added to a plate, followed by 25 μL of the detection antibody mixture, and then the plate was shaken at 705 rpm at room temperature for 2 hours. 100 μL of ECL reading buffer was added to the plate. The plate was then read using an ECL reader.

[0257] Figure 4D shows a "mock ECL-labeled" assay in which a capture antibody on a surface is brought into contact with the analyte mixture, the surface is washed, a detection antibody mixture is added, and the surface is optionally washed again, resulting in a conjugated complex as shown in Figure 4A. Next, an ECL reading buffer is added to the surface along with a detection antibody containing the ECL label but not binding to any component of the conjugated complex on the surface, which acts as a proxy for the "free" ECL label in solution. The resulting ECL is then read by an ECL reading instrument. Figure 4H shows a multiplexed version of the "mock ECL-labeled" assay in which one or more surfaces contain multiple binding domains, each binding domain containing a capture antibody that can bind to the analyte in the analyte mixture. When the surface containing the binding reagent is brought into contact with the analyte mixture, the surface is washed, a detection antibody mixture is added, and the surface is optionally washed again, multiple conjugated complexes are obtained as shown in Figure 4E. Next, an ECL reading buffer is added to the surface along with a detection antibody that contains the ECL label but does not bind to any component of the conjugated complex on the surface. This buffer acts as a proxy for the "free" ECL label in the solution. Then, the generated ECL is read by an ECL reading instrument.

[0258] In the examples herein using a...

Claims

1. (a) 1000 mM to 6500 mM triethanolamine (TEA) and (b) Salt and (c) A composition comprising an electrochemiluminescence (ECL) labeled component, A composition having a pH of 7.0 to 8.0, and substantially free of additional pH buffering components.

2. (a) 1000 mM to 6500 mM triethanolamine (TEA), (b) salt, and (c) surfactant, (a) 1000 mM to 6500 mM triethanolamine (TEA), (b) salt, (c) surfactant, and (d) ECL-labeled component. A composition which is essentially derived from, A composition having a pH of 7.0 to 8.0, and substantially free of additional pH buffering components.

3. The composition according to claim 1 or 2, wherein the composition comprises 500 mM to 2000 mM of the salt.

4. The composition according to claim 1 or 2, wherein the salt contains chloride ions.

5. The composition according to claim 4, wherein the salt comprises NaCl, KCl, LiCl, or a combination thereof.

6. The composition according to claim 1 or 2, wherein the composition comprises an ECL-labeled component, the ECL-labeled component comprises a detection reagent comprising an ECL label, or the ECL-labeled component comprises a binding partner for the detection reagent, the binding partner comprising an ECL label.

7. The composition according to claim 1 or 2, wherein the composition is in a dry state.

8. A method for generating electrochemiluminescence (ECL), (a) The electrodes, (i) A composition comprising 1000 mM to 6500 mM triethanolamine (TEA) and a salt, and (ii) ECL label; To bring it into contact with, (b) A method comprising applying a voltage to the electrode to generate an ECL.

9. A method for detecting bound complexes, (a) Bringing a liquid sample into contact with a surface, wherein the surface is The composition comprises 1000 mM to 6500 mM triethanolamine (TEA) and a salt, The liquid sample contains an ECL-labeled component, or the liquid sample contains a binding partner for the ECL-labeled component, and the method brings the surface into contact with the ECL-labeled component. This further includes contacting the surface, which involves forming a bound complex containing the ECL-labeled component on the surface. (b) Applying a voltage to the surface to generate ECL, (c) A method comprising detecting the generated ECL and thereby detecting the binding complex.

10. A method for detecting bound complexes, (a) Forming a binding complex on the surface, wherein the binding complex contains an ECL-labeled component. (b) The bonded complex, Contact with a composition containing 1000 mM to 6500 mM triethanolamine (TEA) and its salt, (c) Applying a voltage to the surface to generate ECL, (d) A method comprising detecting the generated ECL and thereby detecting the bound complex.

11. A method for detecting a target analyte in a sample, (a) Contacting the sample with (i) a surface containing a binding reagent, wherein the binding reagent specifically binds to the analyte, and (ii) a detection reagent specifically binding to the analyte, wherein the detection reagent contains an ECL label, thereby forming a binding complex on the surface comprising the binding reagent, the analyte, and the detection reagent. (b) The bonded complex on the surface, Contact with a composition containing 1000 mM to 6500 mM triethanolamine (TEA) and its salt, (c) Applying a voltage to the surface to generate ECL, (d) A method comprising detecting the generated ECL and thereby detecting the analyte.

12. The method according to any one of claims 8 to 11, wherein the method does not include a washing step.

13. A method for detecting bound complexes, (a) The sample, i. A composition comprising 1000 mM to 6500 mM triethanolamine (TEA) and a salt, and ii. A detection mixture comprising at least two copies of a detection reagent, wherein each copy of the detection reagent, comprising an ECL label, is combined with the detection mixture to form an assay mixture. (b) The assay mixture, with a binding reagent immobilized on a surface including an electrode, i. A binding complex is formed on the surface, and the binding complex comprises the binding reagent and a first copy of the detection reagent, ii. Contact under conditions that a second copy of the detection reagent remains in the solution, (c) Applying a voltage to the surface to generate ECL, (d) A method comprising detecting the generated ECL and thereby detecting the bound complex.

14. A method for detecting bound complexes, (a) an assay mixture, i. A binding reagent immobilized on the surface, ii. A detection mixture comprising at least two copies of a detection reagent, wherein each copy of the detection reagent comprises an ECL label, and an assay mixture comprising: i. A binding complex is formed on the surface, and the binding complex comprises the binding reagent and a first copy of the detection reagent, ii. Incubating under conditions in which a second copy of the detection reagent remains in the solution, (b) The bonded complex, Contact with a composition containing 1000 mM to 6500 mM triethanolamine (TEA) and its salt, (c) Applying a voltage to the surface to generate ECL, (d) A method comprising detecting the generated ECL and thereby detecting the binding complex.

15. A method for detecting bound complexes, (a) an assay mixture, i. A binding reagent immobilized on the surface, ii. A detection mixture comprising at least two copies of a detection reagent, wherein each copy of the detection reagent comprises an ECL label, iii. An assay mixture comprising a composition containing 1000 mM to 6500 mM triethanolamine (TEA) and its salt, i. A binding complex is formed on the surface, and the binding complex comprises the binding reagent and a first copy of the detection reagent, ii. Incubating under conditions in which a second copy of the detection reagent remains in the solution, (b) Applying a voltage to the surface to generate ECL, (c) A method comprising detecting the generated ECL and thereby detecting the binding complex.

16. A method for quantifying the amount of ECL label in a sample, (a) The electrodes, (i) A composition comprising 1000 mM to 6500 mM triethanolamine (TEA) and a salt, and (ii) Contacting the sample containing the ECL label, (b) Applying a voltage to the electrode, (c) To generate ECL, (d) Measuring the ECL, (e) A method comprising quantifying the amount of the ECL label from the measured ECL.

17. A kit, in which one or more containers, vials, or compartments A kit comprising a composition containing 1000 mM to 6500 mM triethanolamine (TEA) and its salt.

18. The kit according to claim 17, further comprising assay equipment, assay reagents, calibration reagents, ECL labeling, surfaces, or combinations thereof.

19. A composition according to any one of claims 1 to 7, A kit comprising assay equipment, assay reagents, calibration reagents, surfaces, ECL labels, or combinations thereof.

20. The kit according to any one of claims 17 to 19, wherein one or more components of the kit are provided in a dry form.

21. The composition according to claim 1 or 2, wherein the composition has a pH of 7.9.

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