Diagnostic Assay Reagents Comprising Block Copolymers - Patent application

Abstract

The present invention relates to compositions suitable for use in immunoassays and methods of performing immunoassays using the compositions of the invention.The present invention relates to compositions comprising block copolymers and / or chaotropic agents that are useful as buffers, diluents and / or wash solutions in immunoassays and immunoassay methods.

Description

[Technical field]

[0001] The present invention relates to compositions suitable for use in immunoassays and methods of performing immunoassays using the compositions of the invention.The present invention relates to compositions comprising block copolymers and / or chaotropic agents that are useful as buffers, diluents and / or wash solutions in immunoassays and immunoassay methods. [Background technology]

[0002] Diagnostic tests such as immunoassays are often used to detect specific analytes in a sample. For example, pairs of antibodies that can bind to the analyte to form a sandwich that can be detected by an enzyme or label on one or more of the antibodies are well known and are available for a wide range of different analytes of interest. Antibodies against specific biomarkers such as testosterone or cortisol can be used to examine the levels of these substances in saliva, blood or urine samples.

[0003] The presence of the analyte is then determined, for example, using electrochemical or optical measurements. Many electrochemical measurement techniques are known to those skilled in the art, such as electrochemical impedance spectroscopy, differential pulse voltammetry, square wave voltammetry, cyclic voltammetry, chronoamperometry, open circuit potential measurement and chronopotentiometry. Optical detection methods can include fluorescence, chemiluminescence, absorbance and surface plasmon resonance. Other suitable detection techniques can be used to analyze the experimental results in a sensitive and scalable manner. The most common methods are optical detection techniques, electrochemical detection, mechanical analysis, spectroscopy (Raman spectroscopy, NMR spectroscopy) and mass spectrometry (MS). All such techniques can be combined with microfluidic devices.

[0004] Point-of-care detection brings diagnostic testing conveniently and quickly to the subject, allowing better and faster clinical decisions to be made. However, integrating diagnostic tests into point-of-care devices or systems is challenging. Preparation of samples for immunoassays may require mixing multiple solutions and compositions with precise control of volumes and mixing times. Point-of-care systems also require very fast binding kinetics to allow for the typical short test times (<15 min). Furthermore, the device would ideally be automated to eliminate the need for a medical professional to be present.

[0005] Existing liquid handling devices typically flow multiple liquids (e.g., sample liquids, compositions or wash buffers) in the same flow direction across a measurement chamber, reaction zone or other detection means (i.e., different liquids flow sequentially through the same conduits and portions of the device). Alternatively, suitable compositions can be used in traditional laboratory-based devices such as multi-well plates (e.g., 384-well or 96-well plates).

[0006] Current compositions used in diagnostic immunoassays can be subject to matrix effects or non-specific reactions and / or can generate significant background signals, thereby affecting the sensitivity of the assay. Summary of the Invention [Problem to be solved by the invention]

[0007] Thus, there is a need to provide improved diagnostic assay compositions for use in point-of-care immunoassay diagnostic testing, specifically stable compositions that minimize background signal, reduce variability between biological samples, and / or improve the stability, precision, and reproducibility of the immunoassay. [Means for solving the problem]

[0008] This summary introduces concepts that are described in more detail in the detailed description, and is not intended to identify essential features of the claimed subject matter, nor should it be used to limit the scope of the claimed subject matter.

[0009] The present invention provides a composition for an immunoassay comprising a buffer compound, a salt and one or more block copolymers.

[0010] In some embodiments, the buffering compound is selected from the group consisting of Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), Bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), Tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid), is selected from the list consisting of HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), cacodylic acid (dimethylarsenic acid) and MES (2-(N-morpholino)ethanesulfonic acid).

[0011] In some embodiments, the salt is sodium chloride, aluminum ammonium sulfate, aluminum chloride, aluminum chloride, aluminum fluoride, aluminum nitrate, aluminum potassium sulfate, aluminum sulfate, antimony (III) sulfide, antimony trichloride, barium bromide, barium carbonate, barium chloride, barium chromate, barium nitrate, barium perchlorate, barium sulfate, bismuth carbonate, bismuth nitrate, bismuth oxychloride, bismuth subnitrate, bismuth sulfate, bismuth trichloride, cadmium carbonate, cadmium chloride, cadmium iodide, cadmium nitrate, cadmium carbonate, cadmium chloride, cadmium iodide, cadmium nitrate, cadmium carbonate, cadmium chloride, cadmium iodide, cadmium nitrate, cadmium nitrate, cadmium oxychloride, bismuth subnitrate, bismuth sulfate, bismuth trichloride, cadmium carbonate, cadmium chloride, cadmium iodide, cadmium nitrate ... cadmium, cadmium oxide, cadmium sulfate, calcium bromide, calcium carbonate, calcium chloride, calcium fluoride, calcium nitrate, calcium oxide powder, calcium sulfate, chromium (III) chloride, chromium (III) nitrate, chromium (III) oxide, chromium piconylate, chromium trioxide, cobalt (II) carbonate, cobalt (II) chloride, cobalt (II) nitrate, cobalt (II) sulfate, cobalt oxide, cupric bromide, cupric carbonate, cupric chloride, cupric nitrate, cupric oxide powder, copper sulfate, cuprous bromide, cuprous chloride, copper (I) iodide, copper (I) oxide, dysprosium oxide, copper oxide Europium, ferric chloride, ferric nitrate, ferric oxide, ferric sulfate, ferric chloride, ferrous sulfate, gadolinium (III) oxide, germanium dioxide, holmium oxide, indium (III) chloride, iridium trichloride, lanthanum carbonate, lanthanum chloride, lanthanum nitrate, lanthanum oxalate, lanthanum oxide, lead (II) chloride, lead acetate, lead bromide, lead carbonate, lead dioxide, lead monoxide, lead nitrate, lead oxide, lead sulfate, magnesium bromide, magnesium nitrate, magnesium oxide, manganese (II) sulfate, manganese carbonate, manganese dioxide, mercury iodide, mercury nitrate, mercury oxide, molybdenum trioxide , neodymium oxide, nickel carbonate, nickel chloride, nickel nitrate, nickel oxide, nickel sulfate, niobium pentoxide, platinum chloride, potassium bromide, potassium carbonate, potassium chloride, potassium chloroplatinate, potassium chromate, potassium iodate, potassium iodide, potassium nitrate, potassium pyroantimonate, rhodium trichloride, rubidium chloride, samarium oxide, selenium dioxide, silver benzoate, silver bromide, silver carbonate, silver chloride, silver iodide, silver nitrate, silver oxide, silver sulfate, sodium arsenate, sodium arsenite, sodium bromide, sodium carbonate, sodium fluoride, sodium iodide,The fluoride is selected from the list consisting of sodium metaborate, sodium oxalate, sodium silicate, sodium silicofluoride, sodium stannate, sodium tellurite, stannous chloride stannate, stannic oxide, stannous chloride, stannous sulfate, strontium carbonate, titanium tetrachloride, tungsten(VI) oxide, vanadium pentoxide, yttrium oxide, zinc chloride, zinc cyanide, zinc nitrate, zinc oxide, zinc sulfate, zirconium dioxide and zirconyl nitrate.

[0012] In some embodiments, the buffer compound is present at a concentration of about 1 mM to about 500 mM, e.g., about 1 mM to about 450 mM, about 1 mM to about 400 mM, about 1 mM to about 350 mM, about 1 mM to about 300 mM, about 1 mM to about 250 mM, about 1 mM to about 200 mM, about 1 mM to about 150 mM, about 1 mM to about 100 mM, about 1 mM to about 90 mM, about 1 mM to about 80 mM, about 1 mM to about 70 mM, about 1 mM to about 60 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 1 mM to about 10 mM, about 10 mM to about 500 mM, about 10 mM M ~ about 450mM, about 10mM - about 400mM, about 10mM - about 350mM, about 10mM - about 300mM, about 10mM - about 250mM, about 10mM to about 200mM, about 10mM to about 150mM, about 10mM to about 100mM, about 10mM to about 90mM, about 10mM to about 80mM, about 10mM to about 70mM, about 10mM to about 60mM, about 10mM to about 50mM, about 10mM to about 40mM, about 10mM to about 30mM, about 10 mM~about 20mM, about 20mM~about 500mM, about 20mM~about 450mM, about 20mM~about 400mM, about 20mM~about 350mM, about 2 0mM to about 300mM, about 20mM to about 250mM, about 20mM to about 200mM, about 20mM to about 150mM, about 20mM to about 100mM , about 20mM to about 90mM, about 20mM to about 80mM, about 20mM to about 70mM, about 20mM to about 60mM, about 20mM to about 50mM, about 2 0mM to about 40mM, about 20mM to about 30mM, about 30mM to about 500mM, about 30mM to about 450mM, about 30mM to about 400mM, about 30mM to about 350mM, about 30mM to about 300mM, about 30mM to about 250mM, about 30mM to about 200mM, about 30mM to about 150mM , about 30mM to about 100mM, about 30mM to about 90mM, about 30mM to about 80mM, about 30mM to about 70mM, about 30mM to about 60mM, Approximately 30mM to approximately 50mM, approximately 30mM to approximately 40mM, approximately 40mM to approximately 500mM, approximately 40mM to approximately 450mM, approximately 40mM to approximately 400mM, About 40 mM to about 350 mM, about 40 mM to about 300 mM, about 40 mM to about 250 mM, about 40 mM to about 200 mM, about 40 mM to about 150 mM, about 40 mM to about 100 mM, about 40 mM to about 90 mM, about 40 mM to about 80 mM, about 40 mM to about 70 mM, about 40 mM to about 60 mM,about 40mM to about 50mM, about 50mM to about 500mM, about 50mM to about 450mM, about 50mM to about 400mM, about 50mM to about 350mM, about 50mM to about 300mM, about 50mM to about 250mM, about 50mM to about 200mM, about 50mM to about 150mM, about 50mM to about 100mM M, about 50mM to about 90mM, about 50mM to about 80mM, about 50mM to about 70mM, about 50mM to about 60mM, about 60mM to about 500mM, about 60mM to about 450mM, about 60mM to about 400mM, about 60mM to about 350mM, about 60mM to about 300mM, about 60mM to about 250mM , about 60mM to about 200mM, about 60mM to about 150mM, about 60mM to about 100mM, about 60mM to about 90mM, about 60mM to about 80mM, about 60mM to about 70mM, about 70mM to about 500mM, about 70mM to about 450mM, about 70mM to about 400mM, about 70mM to about 350mM , about 70mM to about 300mM, about 70mM to about 250mM, about 70mM to about 200mM, about 70mM to about 150mM, about 70mM to about 100mM, about 70mM to about 90mM, about 70mM to about 80mM, about 80mM to about 500mM, about 80mM to about 450mM, about 80mM to about 400mM, about 80mM to about 350mM, about 80mM to about 300mM, about 80mM to about 250mM, about 80mM to about 200mM, about 80mM to about 150mM, about 80mM to about 100mM, about 80mM to about 90mM, about 90mM to about 500mM, about 9 0mM to about 450mM, about 90mM to about 400mM, about 90mM to about 350mM, about 90mM to about 300mM, about 90mM to about 250mM, about 90mM to about 200mM, about 90mM to about 150mM, about 90mM to about 100mM, about 100mM to about 500mM, about 100mM to about 450mM mM, about 100 mM to about 400 mM, about 100 mM to about 350 mM, about 100 mM to about 300 mM, about 100 mM to about 250 mM, about 100 mM to about 200 mM, about 100 mM to about 150 mM, about 150 mM to about 500 mM, about 150 mM to about 450 mM, about 150 mM to about 400 mM M, about 150mM to about 350mM, about 150mM to about 300mM, about 150mM to about 250mM, about 150mM to about 200mM, about 200mM to about 500mM, about 200mM to about 450mM, about 200mM to about 400mM, about 200mM to about 350mM, about 200mM to about 300mM,The concentration is about 200 mM to about 250 mM, about 250 mM to about 500 mM, about 250 mM to about 450 mM, about 250 mM to about 400 mM, about 250 mM to about 350 mM, about 250 mM to about 300 mM, about 300 mM to about 500 mM, about 300 mM to about 450 mM, about 300 mM to about 400 mM, about 300 mM to about 350 mM, about 350 mM to about 500 mM, about 350 mM to about 450 mM, about 350 mM to about 400 mM, about 400 mM to about 500 mM, about 400 mM to about 450 mM, or about 450 mM to about 500 mM.

[0013] In some embodiments, the composition is present at a concentration of about 1 mM to about 100 mM, for example, about 1 mM to about 90 mM, about 1 mM to about 80 mM, about 1 mM to about 70 mM, about 1 mM to about 60 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 1 mM to about 10 mM, about 10 mM to about 100 mM, about 10 mM to about 90 mM, about 10 mM to about 80 mM, about 10 mM to about 70 mM, about 10 mM to about 100 mM, mM to about 60mM, about 10mM to about 50mM, about 10mM to about 40mM, about 10mM to about 30mM, about 10mM to about 20mM, about 20mM to about 100mM, about 20mM to about 90mM, about 20mM to about 80mM, Approximately 20mM to approximately 70mM, approximately 20mM to approximately 60mM, approximately 20mM to approximately 50mM, approximately 20mM to approximately 40mM, approximately 20mM to approximately 30mM, approximately 30mM to approximately 100mM, approximately 30mM to approximately 90mM, approximately 30mM to approximately 80 mM, about 30mM to about 70mM, about 30mM to about 60mM, about 30mM to about 50mM, about 30mM to about 40mM, about 40mM to about 100mM, about 40mM to about 90mM, about 40mM to about 80mM, about 40mM to Approximately 70mM, approximately 40mM to approximately 60mM, approximately 40mM to approximately 50mM, approximately 50mM to approximately 100mM, approximately 50mM to approximately 90mM, approximately 50mM to approximately 80mM, approximately 50mM to approximately 70mM, approximately 50mM to approximately 60mM, approximately 60 Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 100 mM, about 60 mM to about 90 mM, about 60 mM to about 80 mM, about 60 mM to about 70 mM, about 70 mM to about 100 mM, about 70 mM to about 90 mM, about 70 mM to about 80 mM, about 80 mM to about 100 mM, about 80 mM to about 90 mM, or about 90 mM to about 100 mM.

[0014] In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 10 mM to 100 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to 30 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM.

[0015] In some embodiments, the salt is present at a concentration of about 1 mM to about 1000 mM, for example, about 1 mM to about 900 mM, about 1 mM to about 800 mM, about 1 mM to about 700 mM, about 1 mM to about 600 mM, about 1 mM to about 500 mM, about 1 mM to about 400 mM, about 1 mM to about 300 mM, about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 1 mM to about 50 mM, about 50 mM to about 1000 mM, about 50 mM to about 900 mM, about 50 mM to about 800 mM, about 50 mM to about 700 mM, about 50 mM to about 600 mM, about 50 mM to about 500 mM, about 50 mM to about 400 mM, about 50 mM to about 300 mM, about 50 mM to about 200 mM, about 50 mM to about 100 mM, about 100 mM to about 1000 mM, about 100 mM to about 900 mM, about 100 mM to about 800 mM, about 100 mM to about 700 mM, about 100 mM to about 600 mM, about 100 mM to about 500 mM, about 100 mM to about 400 mM, about 100 mM to about 300 mM, about 100 mM to about 200 mM, about 200 mM to about 1000 mM, about 200 mM to about 900 mM, about 200 mM to about 800 mM, about 200 mM to about 700 mM, about 200mM to about 600mM, about 200mM to about 500mM, about 200mM to about 400mM, about 200mM to about 300mM, about 300mM to about 1000mM, about 300mM to about 900mM, about 300mM to about 800mM, about 300mM to about 700mM, Approximately 300mM to approximately 600mM, approximately 300mM to approximately 500mM, approximately 300mM to approximately 400mM, approximately 400mM to approximately 1000mM, approximately 400mM to about 900mM, about 400mM to about 800mM, about 400mM to about 700mM, about 400mM to about 600mM, about 400 The concentration is from about 500 mM to about 500 mM, from about 500 mM to about 1000 mM, from about 500 mM to about 900 mM, from about 500 mM to about 800 mM, from about 500 mM to about 700 mM, from about 500 mM to about 600 mM, from about 600 mM to about 1000 mM, from about 600 mM to about 900 mM, from about 600 mM to about 800 mM, from about 600 mM to about 700 mM, from about 700 mM to about 1000 mM, from about 700 mM to about 900 mM, from about 700 mM to about 800 mM, from about 800 mM to about 1000 mM, from about 800 mM to about 900 mM, or from about 900 mM to about 1000 mM.

[0016] In some embodiments, the composition is present at a concentration of about 1 mM to about 1000 mM, for example, about 1 mM to about 900 mM, about 1 mM to about 800 mM, about 1 mM to about 700 mM, about 1 mM to about 600 mM, about 1 mM to about 500 mM, about 1 mM to about 400 mM, about 1 mM to about 300 mM, about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 1 mM to about 50 mM, about 50 mM to about 1000 mM, about 50 mM to about 900 mM, about 50 mM to about 800 mM, about 50 mM to about 700 mM, about 50 mM to about 600 mM, about 50 mM to about 500 mM, 50 mM to about 400 mM, about 50 mM to about 300 mM, about 50 mM to about 200 mM, about 50 mM to about 100 mM, about 100 mM to about 1000 mM, about 100 mM to about 900 mM, about 100 mM to about 800 mM, about 100 mM to about 700 mM, about 100 mM to about 600 mM, about 100 mM to about 500 mM, about 100 mM to about 400 mM, about 100 mM to about 300 mM, about 100 mM to about 200 mM, about 200 mM to about 1000 mM, about 200 mM to about 900 mM, about 200 mM to about 800 mM, about 200 mM to about 700 mM, about 00mM to about 600mM, about 200mM to about 500mM, about 200mM to about 400mM, about 200mM to about 300mM, about 300mM to about 1000mM, about 300mM to about 900mM, about 300mM to about 800mM, about 300mM to about 700mM, about 300mM to about 600mM, about 300mM to about 500mM, about 300mM to about 400mM, about 400mM to about 1000mM, about 400mM to about 900mM, about 400mM to about 800mM, about 400mM to about 700mM, about 400mM to about 600mM, about 400mM to about 500mM , about 500 mM to about 1000 mM, about 500 mM to about 900 mM, about 500 mM to about 800 mM, about 500 mM to about 700 mM, about 500 mM to about 600 mM, about 600 mM to about 1000 mM, about 600 mM to about 900 mM, about 600 mM to about 800 mM, about 600 mM to about 700 mM, about 700 mM to about 1000 mM, about 700 mM to about 900 mM, about 700 mM to about 800 mM, about 800 mM to about 1000 mM, about 800 mM to about 900 mM, or about 900 mM to about 1000 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 50 to about 500 mM.In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 200 to about 400 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 100 to about 200 mM.

[0017] In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of 150 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of 400 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 275 mM.

[0018] In some embodiments, the compositions comprise tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), sodium chloride, and one or more block copolymers.

[0019] In some embodiments, the composition comprises Tris-buffered saline (TBS). In some embodiments, the Tris-buffered saline (TBS) comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to 30 mM and NaCl at a concentration of about 100 mM to 200 mM. In some embodiments, the Tris-buffered saline (TBS) comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM and NaCl at a concentration of 150 mM. In some embodiments, the Tris-buffered saline (TBS) contains Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to 30 mM and NaCl at a concentration of about 50 mM to 500 mM. In some embodiments, the Tris-buffered saline (TBS) contains Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM and NaCl at a concentration of 400 mM.

[0020] In some embodiments, the composition comprises Tris-Buffered Saline (TBS) containing Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) and NaCl, and additional NaCl. The NaCl from the TBS is combined with the additional NaCl to obtain the NaCl concentration in the composition. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 200 mM to about 500 mM. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 200 mM to about 400 mM. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a concentration of about 275 mM.

[0021] In some embodiments, the composition is present at a total concentration of about 0.01% (w / v) to about 2% (w / v), e.g., about 0.01% (w / v) to about 1.9% (w / v), about 0.01% (w / v) to about 1.8% (w / v), about 0.01% (w / v) to about 1.7% (w / v), about 0.01% (w / v) to about 1.6% (w / v), about 0.01% (w / v) to about 1.5% (w / v), about 0.01% (w / v) to about 1.4% (w / v), about 0.01% (w / v) to about 1.3% (w / v), about 0.01% (w / v) to about 1.2% (w / v), about 0.01% (w / v) to about 1.1% (w / v), or any combination thereof. w / v), approx. 0.01% (w / v) ~ approx. 1% (w / v), approx. 0.01% (w / v) ~ approx. 0.9% (w / v), approx. 0.01% (w / v) ~ approx. 0.8% (w / v), approximately 0.01% (w / v) ~ approximately 0.7% (w / v), approximately 0.01% (w / v) ~ approximately 0.6% (w / v), approximately 0.01% (w / v) ~ approx. 0.5% (w / v), approx. 0.01% (w / v) ~ approx. 0.4% (w / v), approx. 0.01% (w / v) ~ approx. 0.3% (w / v) , about 0.01% (w / v) to about 0.2% (w / v), about 0.01% (w / v) to about 0.1% (w / v), about 0.01% (w / v) to about 0.0 5% (w / v), approximately 0.05% (w / v) ~ approximately 2% (w / v), approximately 0.05% (w / v) ~ approximately 1.9% (w / v), approximately 0.05% (w / v) ) ~ approx. 1.8% (w / v), approx. 0.05% (w / v) ~ approx. 1.7% (w / v), approx. 0.05% (w / v) ~ approx. 1.6% (w / v), approx. 0. 05% (w / v) ~ approx. 1.5% (w / v), approx. 0.05% (w / v) ~ approx. 1.4% (w / v), approx. 0.05% (w / v) ~ approx. 1.3% (w / v), approx. 0.05% (w / v) ~ approx. 1.2% (w / v), approx. 0.05% (w / v) ~ approx. 1.1% (w / v), approx. 0.05% (w / v) ~ approx. 1% (w / v), approximately 0.05% (w / v) ~ approximately 0.9% (w / v), approximately 0.05% (w / v) ~ approximately 0.8% (w / v), approximately 0.05% (w / v) ~ approx. 0.7% (w / v), approx. 0.05% (w / v) ~ approx. 0.6% (w / v), approx. 0.05% (w / v) ~ approx. 0.5% (w / v), approx. 0.05% (w / v) ~ approx. 0.4% (w / v), approx. 0.05% (w / v) ~ approx. 0.3% (w / v), approx. 0.05% (w / v) ~ approx. 0.2% (w / v), about 0.05% (w / v) to about 0.1% (w / v), about 0.1% (w / v) to about 2% (w / v), about 0.1% (w / v) to about 1.9% (w / v), about 0.1% (w / v) to about 1.8% (w / v), about 0.1% (w / v) to about 1.7% (w / v), about 0.1% (w / v) to about 1.6% (w / v), about 0.1% (w / v) to about 1.5% (w / v), about 0.1% (w / v) to about 1.4% (w / v), about 0.1% (w / v) to about 1.3% (w / v), about 0.1% (w / v) to about 1.2% (w / v), about 0.1% (w / v) to about 1.1% (w / v), about 0.1% (w / v) to about 1% (w / v), about 0.1% (w / v) to about 0.9% (w / v), about 0.1% (w / v) to about 0.8% (w / v), About 0.1% (w / v) to about 0.7% (w / v), about 0.1% (w / v) to about 0.6% (w / v), about 0.1% (w / v) to about 0.5% (w / v), about 0.1% (w / v) to about 0.4% (w / v), about 0.1% (w / v) to about 0.3% (w / v), about 0.1% (w / v) to about 0.2% (w / v), about 0.2% (w / v) to about 2% (w / v), about 0.2% (w / v) to about 1.9% (w / v), about 0.2% (w / v) to about 1.8% (w / v), about 0.2% (w / v) to about 1.7% (w / v), about 0.2% (w / v) to about 1.6% (w / v), about 0.2% (w / v) v) to about 1.5% (w / v), about 0.2% (w / v) to about 1.4% (w / v), about 0.2% (w / v) to about 1.3% (w / v), about 0.2% (w / v) to about 1.2% (w / v), about 0.2% (w / v) to about 1.1% (w / v), about 0.2% (w / v) to about 1% (w / v), about 0.2% (w / v) to about 0.9% (w / v), about 0.2% (w / v) to about 0.8% (w / v), about 0.2% (w / v) to about 0.7% (w / v), about 0.2% (w / v) to about 0.6% (w / v), about 0.2% (w / v) to about 0.5% (w / v), about 0.2% (w / v) to about 0.4% (w / v), about 0.2% (w / v) to about 0.3% (w / v), about 0.3% (w / v) to about 2% (w / v), about 0.3% (w / v) to about 1.9% (w / v), about 0.3% (w / v) to about 1.8% (w / v), about 0.3% (w / v) to about 1.7% (w / v), about 0.3% (w / v) to about 1.6% (w / v), about 0.3% (w / v) to about 1.5% (w / v), about 0.3% (w / v) to about 1.4% (w / v), about 0.3% (w / v) to about 1.3% (w / v), about 0.3% (w / v) to about 1.2% (w / v), about 0.3% (w / v) to about 1.1% (w / v), about 0.3% (w / v) to about 1% (w / v), about 0.3% (w / v) to about 0.9% (w / v), about 0.3% (w / v) to about 0.8% (w / v), about 0.3% (w / v) to about 0.7% (w / v), about 0.3% (w / v) to about 0.6% (w / v), about 0.3% (w / v) to about 0.5% (w / v), about 0.3% (w / v) to about 0.4% (w / v), about 0.4% (w / v) to about 2% (w / v), about 0.4% (w / v) to about 1.9% (w / v), about 0.4% (w / v) to about 1.8% (w / v), about 0.4% (w / v) to about 1.7% (w / v), about 0.4% (w / v) to about 1.6 % (w / v), about 0.4% (w / v) to about 1.5% (w / v), about 0.4% (w / v) to about 1.4% (w / v), about 0.4% (w / v) to about 1.3% (w / v), about 0.4% (w / v) to about 1.2% (w / v), about 0.4% (w / v) to about 1.1% (w / v), about 0.4% (w / v) to about 1% (w / v), about 0.4% (w / v) to about 0.9% (w / v), about 0.4% (w / v) to about 0.8% (w / v), about 0.4% (w / v) to about 0.7% (w / v), about 0.4% (w / v) to about 0.6% (w / v), about 0.4% (w / v) to about 0.5% (w / v), about 0. 5% (w / v) to about 2% (w / v), about 0.5% (w / v) to about 1.9% (w / v), about 0.5% (w / v) to about 1.8% (w / v), about 0.5% (w / v) to about 1.7% (w / v), about 0.5% (w / v) to about 1.6% (w / v), about 0.5% (w / v) to about 1.5% (w / v), about 0.5% (w / v) to about 1.4% (w / v), about 0.5% (w / v) to about 1.3% (w / v), about 0.5% (w / v) to about 1.2% (w / v), about 0.5% (w / v) to about 1.1% (w / v), about 0.5% (w / v) to about 1% (w / v), about 0.5% (w / v) to about 0.9 % (w / v), about 0.5% (w / v) to about 0.8% (w / v), about 0.5% (w / v) to about 0.7% (w / v), about 0.5% (w / v) to about 0.6% (w / v), about 0.6% (w / v) to about 2% (w / v), about 0.6% (w / v) to about 1.9% (w / v), about 0.6% (w / v) to about 1.8% (w / v), about 0.6% (w / v) to about 1.7% (w / v), about 0.6% (w / v) to about 1.6% (w / v), about 0.6% (w / v) to about 1.5% (w / v), about 0.6% (w / v) to about 1.4% (w / v), about 0.6% (w / v) to about 1.3% (w / v), about 0.6% (w / v) to about 1.2% (w / v), about 0.6% (w / v) to about 1.1% (w / v), about 0.6% (w / v) to about 1% (w / v), about 0.6% (w / v) to about 0.9% (w / v), about 0.6% (w / v) to about 0.8% (w / v), about 0.6% (w / v) to about 0.7% (w / v), about 0.7% (w / v) to about 2% (w / v), about 0.7% (w / v) to about 1.9% (w / v), about 0.7% (w / v) to about 1.8% (w / v), about 0.7% (w / v) to about 1.7% (w / v), about 0.7% (w / v) to about 1.6% (w / v), about 0.7% (w / v) to about 1. 5% (w / v), about 0.7% (w / v) to about 1.4% (w / v), about 0.7% (w / v) to about 1.3% (w / v), about 0.7% (w / v) to about 1.2% (w / v), about 0.7% (w / v) to about 1.1% (w / v), about 0.7% (w / v) to about 1% (w / v), about 0.7% (w / v) to about 0.9% (w / v), about 0.7% (w / v) to about 0.8% (w / v), about 0.8% (w / v) to about 2% (w / v), about 0.8% (w / v) to about 1.9% (w / v), about 0.8% (w / v) to about 1.8% (w / v), about 0.8% (w / v) to about 1.7% (w / v), about 0. 8% (w / v) to about 1.6% (w / v), about 0.8% (w / v) to about 1.5% (w / v), about 0.8% (w / v) to about 1.4% (w / v), about 0.8% (w / v) to about 1.3% (w / v), about 0.8% (w / v) to about 1.2% (w / v), about 0.8% (w / v) to about 1.1% (w / v), about 0.8% (w / v) to about 1% (w / v), about 0.8% (w / v) to about 0.9% (w / v), about 0.9% (w / v) to about 2% (w / v), about 0.9% (w / v) to about 1.9% (w / v), about 0.9% (w / v) to about 1.8% (w / v), about 0.9% (w / v) to about 1. 7% (w / v), about 0.9% (w / v) to about 1.6% (w / v), about 0.9% (w / v) to about 1.5% (w / v), about 0.9% (w / v) to about 1.4% (w / v), about 0.9% (w / v) to about 1.3% (w / v), about 0.9% (w / v) to about 1.2% (w / v), about 0.9% (w / v) to about 1.1% (w / v), about 0.9% (w / v) to about 1% (w / v), about 1% (w / v) to about 2% (w / v), about 1% (w / v) to about 1.9% (w / v), about 1% (w / v) to about 1.8% (w / v), about 1% (w / v) to about 1.7% (w / v), about 1% (w / v) to about 1.6% (w / v), about 1% (w / v) to about 1.5% (w / v), about 1% (w / v) to about 1.4% (w / v), about 1% (w / v) to about 1.3% (w / v), about 1% (w / v) to about 1.2% (w / v), about 1% (w / v) to about 1.1% (w / v), about 1.1% (w / v) to about 2% (w / v), about 1.1% (w / v) to about 1.9% (w / v), about 1.1% (w / v) to about 1.8% (w / v), about 1.1% (w / v) to about 1.7% (w / v), about 1.1% (w / v) to about 1.6% (w / v), about 1.1% (w / v) to about 1.5% (w / v), about 1.1% (w / v) to about about 1.4% (w / v), about 1.1% (w / v) to about 1.3% (w / v), about 1.1% (w / v) to about 1.2% (w / v), about 1.2% (w / v) to about 2% (w / v), about 1.2% (w / v) to about 1.9% (w / v), about 1.2% (w / v) to about 1.8% (w / v), about 1.2% (w / v) to about 1.7% (w / v), about 1.2% (w / v) to about 1.6% (w / v), about 1.2% (w / v) to about 1.5% (w / v), about 1.2% (w / v) to about 1.4% (w / v), about 1.2% (w / v) to about 1.3% (w / v), about 1.3% (w / v) to about 2% (w / v), about 1.3% (w / v) to about 1.9% (w / v), about 1.3% (w / v) to about 1.8% (w / v), about 1.3% (w / v) to about 1.7% (w / v), about 1.3% (w / v) to about 1.6% (w / v), about 1.3% (w / v) to about 1.5% (w / v), about 1.3% (w / v) to about 1.4% (w / v), about 1.4% (w / v) to about 2% (w / v), about 1.4% (w / v) to about 1.9% (w / v), about 1.4% (w / v) to about 1.8% (w / v), about 1.4% (w / v) to about 1.7% (w / v), about 1.4% (w / v) to about 1.6% (w / v), about 1.4% (w / v) to About 1.5% (w / v), about 1.5% (w / v) to about 2% (w / v), about 1.5% (w / v) to about 1.9% (w / v), about 1.5% (w / v) to about 1.8% (w / v), about 1.5% (w / v) to about 1.7% (w / v), about 1.5% (w / v) to about 1.6% (w / v), about 1.6% (w / v) to about 2% (w / v), about 1.6% (w / v) to about 1.9% (w / v), about 1.6% (w / v) to about 1.8% (w / v), about 1.6% (w / v) to about 1.7% (w / v), about 1.7% (w / v) to about 2% (w / v), about 1.7% (w / v) to about 1.9% (w / v), about 1.6% (w / v) to about 1.8% (w / v), about 1.6% (w / v) to about 1.7% (w / v), about 1.7% (w / v) to about 2% (w / v), about 1.7% (w / v) to about 1.9% (w / v), about 1.The composition comprises one or more block copolymers in a total concentration of from about 7% (w / v) to about 1.8% (w / v), from about 1.8% (w / v) to about 2% (w / v), from about 1.8% (w / v) to about 1.9% (w / v), or from about 1.9% (w / v) to about 2% (w / v).

[0022] In some embodiments, the composition comprises one or more block copolymers at a total concentration of about 0.25% (w / v) to about 1.5% (w / v). In some embodiments, the composition comprises one or more block copolymers at a total concentration of about 0.05% (w / v). In some embodiments, the composition comprises one or more block copolymers at a total concentration of about 0.125% (w / v). In some embodiments, the composition comprises one block copolymer at a total concentration of about 0.05% (w / v). In some embodiments, the composition comprises one block copolymer at a total concentration of about 0.125% (w / v).

[0023] In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to 1000 mM, and one or more block copolymers at a total concentration of about 0.01% (w / v) to 2% (w / v). In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM, sodium chloride (NaCl) at a concentration of 150 mM, and block copolymer at a concentration of 0.05% (w / v). In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM, sodium chloride (NaCl) at a concentration of 275 mM, and a block copolymer at a concentration of 0.125% (w / v).

[0024] The present invention also provides compositions for immunoassays comprising a buffer compound, one or more salts, one or more blocking agents, and one or more block copolymers. In some embodiments, the one or more blocking agents are selected from the list consisting of a protein, such as a phosphoprotein, a polymer, and a small molecule. In some embodiments, the protein is a phosphoprotein, and optionally, the phosphoprotein is casein. In some implementations, the protein is bovine serum albumin (BSA). In some embodiments, the polymer is PVP (polyvinylpyrrolidone) or PVA (polyvinyl alcohol). In some embodiments, the small molecule is ethanolamine.

[0025] In some embodiments, the composition comprises at least two salts, for example, 2, 3, 4, 5 or 6 salts, In some embodiments, the composition comprises two salts.

[0026] In some embodiments, the buffering compound is selected from the group consisting of Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), Bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), Tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropane-1,3-diol), TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropane-1,3-diol), bis(2-(bis(2-hydroxyethyl)amino)acetic acid ... (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), cacodylic acid (dimethylarsenic acid) and MES (2-(N-morpholino)ethanesulfonic acid). In some embodiments, the one or more salts are selected from the group consisting of sodium chloride, calcium chloride, aluminum ammonium sulfate, aluminum chloride, aluminum chloride, aluminum fluoride, aluminum nitrate, aluminum potassium sulfate, aluminum sulfate, antimony (III) sulfide, antimony trichloride, barium bromide, barium carbonate, barium chloride, barium chromate, barium nitrate, barium perchlorate, barium sulfate, bismuth carbonate, bismuth nitrate, bismuth oxychloride, bismuth subnitrate, bismuth sulfate, bismuth trichloride, cadmium carbonate, cadmium chloride, Cadmium iodide, cadmium nitrate, cadmium oxide, cadmium sulfate, calcium bromide, calcium carbonate, calcium fluoride, calcium nitrate, calcium oxide powder, calcium sulfate, chromium(III) chloride, chromium(III) nitrate, chromium(III) oxide, chromium piconylate, chromium trioxide, cobalt(II) carbonate, cobalt(II) chloride, cobalt(II) nitrate, cobalt(II) sulfate, cobalt oxide, cupric bromide, cupric carbonate, cupric chloride, cupric nitrate, cupric oxide powder, copper sulfate, cuprous bromide, cuprous chloride, cuprous iodide, cuprous oxide, copper oxide powder rhodium, europium oxide, ferric chloride, ferric nitrate, ferric oxide, ferric sulfate, ferric chloride, ferrous sulfate, gadolinium(III) oxide, germanium dioxide, holmium oxide, indium(III) chloride, iridium trichloride, lanthanum carbonate, lanthanum chloride, lanthanum nitrate, lanthanum oxalate, lanthanum oxide, lead(II) chloride, lead acetate, lead bromide, lead carbonate, lead dioxide, lead monoxide, lead nitrate, lead oxide, lead sulfate, magnesium bromide, magnesium nitrate, magnesium oxide, manganese(II) sulfate, manganese carbonate, manganese dioxide, mercury iodide, mercury nitrate, mercury oxide , molybdenum trioxide, neodymium oxide, nickel carbonate, nickel chloride, nickel nitrate, nickel oxide, nickel sulfate, niobium pentoxide, platinum chloride, potassium bromide, potassium carbonate, potassium chloride, potassium chloroplatinate, potassium chromate, potassium iodate, potassium iodide, potassium nitrate, potassium pyroantimonate, rhodium trichloride, rubidium chloride, samarium oxide, selenium dioxide, silver benzoate, silver bromide, silver carbonate, silver chloride, silver iodide, silver nitrate, silver oxide, silver sulfate, sodium arsenate, sodium arsenite, sodium bromide, sodium carbonate, sodium fluoride,The fluoride is selected from the list consisting of sodium iodide, sodium metaborate, sodium oxalate, sodium silicate, sodium silicofluoride, sodium stannate, sodium tellurite, stannous chloride stannate, stannic oxide, stannous chloride, stannous sulfate, strontium carbonate, titanium tetrachloride, tungsten(VI) oxide, vanadium pentoxide, yttrium oxide, zinc chloride, zinc cyanide, zinc nitrate, zinc oxide, zinc sulfate, zirconium dioxide and zirconyl nitrate.

[0027] In some embodiments, the buffer compound is present at a concentration of about 1 mM to about 500 mM, e.g., about 1 mM to about 450 mM, about 1 mM to about 400 mM, about 1 mM to about 350 mM, about 1 mM to about 300 mM, about 1 mM to about 250 mM, about 1 mM to about 200 mM, about 1 mM to about 150 mM, about 1 mM to about 100 mM, about 1 mM to about 90 mM, about 1 mM to about 80 mM, about 1 mM to about 70 mM, about 1 mM to about 60 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 1 mM to about 10 mM, about 10 mM to about 500 mM, about 10 mM M ~ about 450mM, about 10mM - about 400mM, about 10mM - about 350mM, about 10mM - about 300mM, about 10mM - about 250mM, about 10mM to about 200mM, about 10mM to about 150mM, about 10mM to about 100mM, about 10mM to about 90mM, about 10mM to about 80mM, about 10mM to about 70mM, about 10mM to about 60mM, about 10mM to about 50mM, about 10mM to about 40mM, about 10mM to about 30mM, about 10 mM~about 20mM, about 20mM~about 500mM, about 20mM~about 450mM, about 20mM~about 400mM, about 20mM~about 350mM, about 2 0mM to about 300mM, about 20mM to about 250mM, about 20mM to about 200mM, about 20mM to about 150mM, about 20mM to about 100mM , about 20mM to about 90mM, about 20mM to about 80mM, about 20mM to about 70mM, about 20mM to about 60mM, about 20mM to about 50mM, about 2 0mM to about 40mM, about 20mM to about 30mM, about 30mM to about 500mM, about 30mM to about 450mM, about 30mM to about 400mM, about 30mM to about 350mM, about 30mM to about 300mM, about 30mM to about 250mM, about 30mM to about 200mM, about 30mM to about 150mM , about 30mM to about 100mM, about 30mM to about 90mM, about 30mM to about 80mM, about 30mM to about 70mM, about 30mM to about 60mM, Approximately 30mM to approximately 50mM, approximately 30mM to approximately 40mM, approximately 40mM to approximately 500mM, approximately 40mM to approximately 450mM, approximately 40mM to approximately 400mM, About 40 mM to about 350 mM, about 40 mM to about 300 mM, about 40 mM to about 250 mM, about 40 mM to about 200 mM, about 40 mM to about 150 mM, about 40 mM to about 100 mM, about 40 mM to about 90 mM, about 40 mM to about 80 mM, about 40 mM to about 70 mM, about 40 mM to about 60 mM,about 40mM to about 50mM, about 50mM to about 500mM, about 50mM to about 450mM, about 50mM to about 400mM, about 50mM to about 350mM, about 50mM to about 300mM, about 50mM to about 250mM, about 50mM to about 200mM, about 50mM to about 150mM, about 50mM to about 100mM M, about 50mM to about 90mM, about 50mM to about 80mM, about 50mM to about 70mM, about 50mM to about 60mM, about 60mM to about 500mM, about 60mM to about 450mM, about 60mM to about 400mM, about 60mM to about 350mM, about 60mM to about 300mM, about 60mM to about 250mM , about 60mM to about 200mM, about 60mM to about 150mM, about 60mM to about 100mM, about 60mM to about 90mM, about 60mM to about 80mM, about 60mM to about 70mM, about 70mM to about 500mM, about 70mM to about 450mM, about 70mM to about 400mM, about 70mM to about 350mM , about 70mM to about 300mM, about 70mM to about 250mM, about 70mM to about 200mM, about 70mM to about 150mM, about 70mM to about 100mM, about 70mM to about 90mM, about 70mM to about 80mM, about 80mM to about 500mM, about 80mM to about 450mM, about 80mM to about 400mM, about 80mM to about 350mM, about 80mM to about 300mM, about 80mM to about 250mM, about 80mM to about 200mM, about 80mM to about 150mM, about 80mM to about 100mM, about 80mM to about 90mM, about 90mM to about 500mM, about 9 0mM to about 450mM, about 90mM to about 400mM, about 90mM to about 350mM, about 90mM to about 300mM, about 90mM to about 250mM, about 90mM to about 200mM, about 90mM to about 150mM, about 90mM to about 100mM, about 100mM to about 500mM, about 100mM to about 450mM mM, about 100 mM to about 400 mM, about 100 mM to about 350 mM, about 100 mM to about 300 mM, about 100 mM to about 250 mM, about 100 mM to about 200 mM, about 100 mM to about 150 mM, about 150 mM to about 500 mM, about 150 mM to about 450 mM, about 150 mM to about 400 mM M, about 150mM to about 350mM, about 150mM to about 300mM, about 150mM to about 250mM, about 150mM to about 200mM, about 200mM to about 500mM, about 200mM to about 450mM, about 200mM to about 400mM, about 200mM to about 350mM, about 200mM to about 300mM,The concentration is about 200 mM to about 250 mM, about 250 mM to about 500 mM, about 250 mM to about 450 mM, about 250 mM to about 400 mM, about 250 mM to about 350 mM, about 250 mM to about 300 mM, about 300 mM to about 500 mM, about 300 mM to about 450 mM, about 300 mM to about 400 mM, about 300 mM to about 350 mM, about 350 mM to about 500 mM, about 350 mM to about 450 mM, about 350 mM to about 400 mM, about 400 mM to about 500 mM, about 400 mM to about 450 mM, or about 450 mM to about 500 mM.

[0028] In some embodiments, the composition is present at a concentration of about 1 mM to about 100 mM, for example, about 1 mM to about 90 mM, about 1 mM to about 80 mM, about 1 mM to about 70 mM, about 1 mM to about 60 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 1 mM to about 10 mM, about 10 mM to about 100 mM, about 10 mM to about 90 mM, about 10 mM to about 80 mM, about 10 mM to about 70 mM, about 10 mM to about 100 mM, mM to about 60mM, about 10mM to about 50mM, about 10mM to about 40mM, about 10mM to about 30mM, about 10mM to about 20mM, about 20mM to about 100mM, about 20mM to about 90mM, about 20mM to about 80mM, Approximately 20mM to approximately 70mM, approximately 20mM to approximately 60mM, approximately 20mM to approximately 50mM, approximately 20mM to approximately 40mM, approximately 20mM to approximately 30mM, approximately 30mM to approximately 100mM, approximately 30mM to approximately 90mM, approximately 30mM to approximately 80 mM, about 30mM to about 70mM, about 30mM to about 60mM, about 30mM to about 50mM, about 30mM to about 40mM, about 40mM to about 100mM, about 40mM to about 90mM, about 40mM to about 80mM, about 40mM to Approximately 70mM, approximately 40mM to approximately 60mM, approximately 40mM to approximately 50mM, approximately 50mM to approximately 100mM, approximately 50mM to approximately 90mM, approximately 50mM to approximately 80mM, approximately 50mM to approximately 70mM, approximately 50mM to approximately 60mM, approximately 60 The composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM, about 60 mM to about 90 mM, about 60 mM to about 80 mM, about 60 mM to about 70 mM, about 70 mM to about 100 mM, about 70 mM to about 90 mM, about 70 mM to about 80 mM, about 80 mM to about 100 mM, about 80 mM to about 90 mM, or about 90 mM to about 100 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM.

[0029] In some embodiments, one of the one or more salts is at a concentration of about 1 mM to 1000 mM, e.g., about 1 mM to about 900 mM, about 1 mM to about 800 mM, about 1 mM to about 700 mM, about 1 mM to about 600 mM, about 1 mM to about 500 mM, about 1 mM to about 400 mM, about 1 mM to about 300 mM, about 1 mM to about 200 mM, or about 200 mM. 0 mM, about 1 mM to about 100 mM, about 10 mM to about 1000 mM, about 10 mM to about 900 mM, about 10 mM to about 800 mM, about 10 mM to about 700 mM, about 10 mM to about 600 mM, about 10 mM to about 500 mM, about 10 mM to about 400 mM, about 10 mM to about 300 mM, about 10 mM to about 200 mM, about 10 mM M ~ about 100mM, about 50mM - about 1000mM, about 50mM - about 900mM, about 50mM - about 800mM, about 50mM - about 700mM, about 50mM - Approximately 600mM, approximately 50mM to approximately 500mM, approximately 50mM to approximately 400mM, approximately 50mM to approximately 300mM, approximately 50mM to approximately 200mM, approximately 50mM to approximately 100 The concentration is about 100 mM to about 1000 mM, about 100 mM to about 900 mM, about 100 mM to about 800 mM, about 100 mM to about 700 mM, about 100 mM to about 600 mM, about 100 mM to about 500 mM, about 100 mM to about 400 mM, about 100 mM to about 300 mM, or about 100 mM to about 200 mM.

[0030] In some embodiments, the composition is present at a concentration of about 1 mM to 1000 mM, for example, about 1 mM to about 900 mM, about 1 mM to about 800 mM, about 1 mM to about 700 mM, about 1 mM to about 600 mM, about 1 mM to about 500 mM, about 1 mM to about 400 mM, about 1 mM to about 300 mM, about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 10 mM to about 1000 mM, about 10 mM to about 900 mM, about 10 mM to about 800 mM, about 10 mM to about 700 mM, about 10 mM to about 600 mM, about 10 mM to about 500 mM, about 10 mM to about 400 mM, about 10 mM to about 300 mM, about 10 mM to about 200 mM, about 10 mM to about 100 mM, about 0 mM to about 1000 mM, about 50 mM to about 900 mM, about 50 mM to about 800 mM, about 50 mM to about 700 mM, about 50 mM to about 600 mM, about 50 mM to about 500 mM, about 50 mM to about 400 mM, about 50 mM to about 300 mM, about 50 mM to about 200 mM, about 50 mM to about 100 mM, about 100 mM to about 10 In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 200 mM to about 400 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 300 mM to about 500 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 100 mM to about 200 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 100 mM to about 200 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of about 400 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of 275 mM. In some embodiments, the composition comprises sodium chloride (NaCl) at a concentration of 150 mM.

[0031] In some embodiments, one of the one or more salts is at a concentration of about 1 mM to about 200 mM, e.g., about 1 mM to about 150 mM, about 1 mM to about 100 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 1 mM to about 10 mM, about 5 mM to about 200 mM, about 5 mM to about 150 mM, about 5 mM to about 100 mM, about 5 mM to about 50 mM, about 5 mM to about 40 mM, about 5 mM to about 30 mM, about 5 mM to about 50 mM, The concentration is about 20 mM, about 5 mM to about 10 mM, about 10 mM to about 200 mM, about 10 mM to about 150 mM, about 10 mM to about 100 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 30 mM, about 10 mM to about 20 mM, about 15 mM to about 200 mM, about 15 mM to about 150 mM, about 15 mM to about 100 mM, about 15 mM to about 50 mM, about 15 mM to about 40 mM, about 15 mM to about 30 mM, or about 15 mM to about 20 mM.

[0032] In some embodiments, the composition is present at a concentration of about 1 mM to about 200 mM, e.g., about 1 mM to about 150 mM, about 1 mM to about 100 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 1 mM to about 10 mM, about 5 mM to about 200 mM, about 5 mM to about 150 mM, about 5 mM to about 100 mM, about 5 mM to about 50 mM, about 5 mM to about 40 mM, about 5 mM to about 30 mM, about 5 mM to about 20 mM, about 5 mM to about Contains calcium chloride (CaCl2) at a concentration of 10 mM, about 10 mM to about 200 mM, about 10 mM to about 150 mM, about 10 mM to about 100 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 30 mM, about 10 mM to about 20 mM, about 15 mM to about 200 mM, about 15 mM to about 150 mM, about 15 mM to about 100 mM, about 15 mM to about 50 mM, about 15 mM to about 40 mM, about 15 mM to about 30 mM, or about 15 mM to about 20 mM.

[0033] In some embodiments, the composition comprises calcium chloride (CaCl2) at a concentration of about 5 mM to 20 mM. In some embodiments, the composition comprises calcium chloride (CaCl2) at a concentration of about 10 mM to 20 mM. In some embodiments, the composition comprises calcium chloride (CaCl2) at a concentration of 16 mM. In some embodiments, the composition comprises calcium chloride (CaCl2) at a concentration of 8 mM. In some embodiments, the composition comprises Tris-Buffered Saline (TBS).

[0034] In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to 30 mM and NaCl at a concentration of about 100 mM to 200 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM and NaCl at a concentration of 150 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 200 mM to 400 mM. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a concentration of about 275 mM.

[0035] In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM, NaCl at a concentration of about 200 mM to about 400 mM, and CaCl2 at a concentration of about 5 mM to about 20 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, NaCl at a concentration of about 275 mM, and CaCl2 at a concentration of about 8 mM.

[0036] In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to 30 mM, NaCl at a concentration of about 300 mM to 500 mM, and CaCl2 at a concentration of about 10 mM to about 20 mM. In some embodiments, the composition comprises Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, NaCl at a concentration of about 400 mM, and CaCl2 at a concentration of about 16 mM.

[0037] In some embodiments, the composition is present at a total concentration of about 0.05% (w / v) to about 5% (w / v), e.g., about 0.05% (w / v) to about 4.5% (w / v), about 0.05% (w / v) to about 4% (w / v), about 0.05% (w / v) to about 3.5% (w / v), about 0.05% (w / v) to about 3% (w / v), about 0.05% (w / v) to about 2.5% (w / v), about 0.05% (w / v) to about 2% (w / v), about 0.05% (w / v) to about 1.5% (w / v), about 0.05% (w / v) to about 1 % (w / v), about 0.05% (w / v) to about 0.5% (w / v), about 0.1% (w / v) to about 5% (w / v), about 0.1% (w / v) to about 4.5% (w / v), about 0.1% (w / v) to about 4% (w / v), about 0.1% (w / v) to about 3.5% (w / v), about 0.1% (w / v) to about 3% (w / v), about 0.1% (w / v) to about 2.5% (w / v), about 0.1% (w / v) to about 2% (w / v), about 0.1% (w / v) to about 1.5% (w / v), about 0.1% (w / v) to about 1% (w / v) ), about 0.1% (w / v) to about 0.5% (w / v), about 0.5% (w / v) to about 5% (w / v), about 0.5% (w / v) to about 4.5% (w / v), about 0.5% (w / v) to about 4% (w / v), about 0.5% (w / v) to about 3.5% (w / v), Approximately 0.5% (w / v) ~ approximately 3% (w / v), approximately 0.5% (w / v) ~ approximately 2.5% (w / v), approximately 0.5% (w / v) ~ approximately 2% (w / v), approximately 0.5% (w / v) ~ approximately 1.5% (w / v), approximately 0.5% (w / v) ~ approximately 1% (w / v), approximately 0.7 The composition comprises one or more block copolymers at a total concentration of about 5% (w / v) to about 5% (w / v), about 0.75% (w / v) to about 4.5% (w / v), about 0.75% (w / v) to about 4% (w / v), about 0.75% (w / v) to about 3.5% (w / v), about 0.75% (w / v) to about 3% (w / v), about 0.75% (w / v) to about 2.5% (w / v), about 0.75% (w / v) to about 2% (w / v), about 0.75% (w / v) to about 1.5% (w / v), or about 0.75% (w / v) to about 1% (w / v). In some embodiments, the composition comprises one or more block copolymers at a total concentration of about 0.75% (w / v) to 1.5% (w / v). In some embodiments, the composition comprises one or more block copolymers at a total concentration of 1% (w / v).In some embodiments, the composition comprises one block copolymer at a concentration of 1% (w / v).

[0038] In some embodiments, the composition comprises one or more block copolymers at a total concentration of about 0.05% (w / v) to 0.5% (w / v). In some embodiments, the composition comprises one or more block copolymers at a total concentration of 0.25% (w / v). In some embodiments, the composition comprises one block copolymer at a concentration of 0.25% (w / v). In some embodiments, the composition comprises one or more block copolymers at a total concentration of 0.125% (w / v). In some embodiments, the composition comprises one block copolymer at a concentration of 0.125% (w / v).

[0039] In some embodiments, the composition is administered at a total concentration of about 0.05% (w / v) to about 10% (w / v), e.g., about 0.05% (w / v) to about 9% (w / v), about 0.05% (w / v) to about 8% (w / v), about 0.05% (w / v) to about 7% (w / v), about 0.05% (w / v) to about 6% (w / v), about 0.05% (w / v) to about 5% (w / v), about 0.05% (w / v) to about 4% (w / v), about 0.05% (w / v) to about 3% (w / v), about 0.05% (w / v) to about 2.5% (w / v), about 0.05% (w / v) to about 2% (w / v), about 0.05% (w / v) ~1.5% (w / v), approx. 0.05% (w / v) ~ approx. 1% (w / v), approx. 0.05% (w / v) ~ approx. 0.5% (w / v), approx. 0.05% (w / v) ~ approx. 0.4% (w / v), approx. 0.05% (w / v) ~ approx. 0.3% (w / v), approx. 0.05% (w / v) ~ approx. 0.2% (w / v), Approx. 0.05% (w / v) ~ approx. 0.1% (w / v), approx. 0.1% (w / v) ~ approx. 10% (w / v), approx. 0.1% (w / v) ~ approx. 9% (w / v) ), about 0.1% (w / v) to about 8% (w / v), about 0.1% (w / v) to about 7% (w / v), about 0.1% (w / v) to about 6% (w / v), about 0.1% (w / v) ~ approx. 5% (w / v), approx. 0.1% (w / v) ~ approx. 4% (w / v), approx. 0.1% (w / v) ~ approx. 3% (w / v), approx. 0. 1% (w / v) ~ approx. 2.5% (w / v), approx. 0.1% (w / v) ~ approx. 2% (w / v), approx. 0.1% (w / v) ~ approx. 1.5% (w / v), approx. 0 .1%(w / v) ~ approx. 1% (w / v), approx. 0.1% (w / v) ~ approx. 0.5% (w / v), approx. 0.1% (w / v) ~ approx. 0.4% (w / v), Approx. 0.1% (w / v) ~ approx. 0.3% (w / v), approx. 0.1% (w / v) ~ approx. 0.2% (w / v), approx. 0.2% (w / v) ~ approx. 10% (w / v) ), about 0.2% (w / v) to about 9% (w / v), about 0.2% (w / v) to about 8% (w / v), about 0.2% (w / v) to about 7% (w / v), Approximately 0.2% (w / v) to approximately 6% (w / v), approximately 0.2% (w / v) to approximately 5% (w / v), approximately 0.2% (w / v) to approximately 4% (w / v), approximately 0. 2% (w / v) ~ approx. 3% (w / v), approx. 0.2% (w / v) ~ approx. 2.5% (w / v), approx. 0.2% (w / v) ~ approx. 2% (w / v), approx. 0.2 %(w / v) ~ approx. 1.5% (w / v), approx. 0.2% (w / v) ~ approx. 1% (w / v), approx. 0.2% (w / v) ~ approx. 0.5% (w / v), approx. 0.2% (w / v) to about 0.4% (w / v), about 0.2% (w / v) to about 0.3% (w / v), about 0.3% (w / v) to about 10% (w / v), about 0.3% (w / v) to about 9% (w / v), about 0.3% (w / v) to about 8% (w / v), about 0.3% (w / v) to about 7% (w / v), about 0.3% (w / v) to about 6% (w / v), about 0.3% (w / v) to about 5% (w / v), about 0.3% (w / v) to about 4% (w / v), about 0.3% (w / v) to about 3% (w / v), about 0.3% (w / v) to about 2.5% (w / v), about 0.3% (w / v) to about 2% (w / v), about 0.3% (w / v) (w / v) to about 1.5% (w / v), about 0.3% (w / v) to about 1% (w / v), about 0.3% (w / v) to about 0.5% (w / v), about 0.3% (w / v) to about 0.4% (w / v), about 0.4% (w / v) to about 10% (w / v), about 0.4% (w / v) to about 9% (w / v), about 0.4% (w / v) to about 8% (w / v), about 0.4% (w / v) to about 7% (w / v), about 0.4% (w / v) to about 6% (w / v), about 0.4% (w / v) to about 5% (w / v), about 0.4% (w / v) to about 4% (w / v), about 0.4% (w / v) to about 3% (w / v), about 0.4% (w / v) v) to about 2.5% (w / v), about 0.4% (w / v) to about 2% (w / v), about 0.4% (w / v) to about 1.5% (w / v), about 0.4% (w / v) to about 1% (w / v), about 0.4% (w / v) to about 0.5% (w / v), about 0.5% (w / v) to about 10% (w / v), about 0.5% (w / v) to about 9% (w / v), about 0.5% (w / v) to about 8% (w / v), about 0.5% (w / v) to about 7% (w / v), about 0.5% (w / v) to about 6% (w / v), about 0.5% (w / v) to about 5% (w / v), about 0.5% (w / v) to about 4% (w / v), about 0.5% (w / v) to about 3% (w / v), about 0.5% (w / v) to about 2.5% (w / v), about 0.5% (w / v) to about 2% (w / v), about 0.5% (w / v) to about 1.5% (w / v), about 0.5% (w / v) to about 1% (w / v), about 1% (w / v) to about 10% (w / v), about 1% (w / v) to about 9% (w / v), about 1% (w / v) to about 8% (w / v), about 1% (w / v) to about 7% (w / v), about 1% (w / v) to about 6% (w / v), about 1% (w / v) to about 5% (w / v), about 1% (w / v) to about 4% (w / v), about 1% (w / v) to about 3% (w / v), about 1% (w / v) to about 2.5% (w / v), about 1% (w / v) to about 2% (w / v), about 1% (w / v) to about 1.5% (w / v), about 1.5% (w / v) to about 10% (w / v), about 1.5% (w / v) to about 9% (w / v), about 1.5% (w / v) to about 8% (w / v), about 1.5% (w / v) to about 7% (w / v), about 1.5% (w / v) to about 6% (w / v), about 1.5% (w / v) to about 5% (w / v), about 1.5% (w / v) to about 4% (w / v), about 1.5% (w / v) to about 3% (w / v), about 1.5% (w / v) to about 2.5% (w / v), about 1.5% (w / v) to about 2% (w / v), about 2% (w / v) to about 10% (w / v), about 2% (w / v) to about 9 % (w / v), about 2% (w / v) to about 8% (w / v), about 2% (w / v) to about 7% (w / v), about 2% (w / v) to about 6% (w / v), about 2% (w / v) to about 5% (w / v), about 2% (w / v) to about 4% (w / v), about 2% (w / v) to about 3% (w / v), about 2% (w / v) to about 2.5% (w / v), about 2.5 % (w / v) to about 10% (w / v), about 2.5% (w / v) to about 9% (w / v), about 2.5% (w / v) to about 8% (w / v), about 2.5% (w / v) to about 7% (w / v), about 2.5% (w / v) to about 6% (w / v), about 2.5% (w / v) to about 5% (w / v), about 2.5% (w / v) to about 4% (w / v), about 2.5% (w / v) ~ approx. 3% (w / v), approx. 3% (w / v) ~ approx. 10% (w / v), approx. 3% (w / v) ~ approx. 9% (w / v), approx. 3% (w / v) ~ approx. 8% (w / v), about 3% (w / v) to about 7% (w / v), about 3% (w / v) to about 6% (w / v), about 3% (w / v) to about 5% (w / v), about 3% (w / v) to about 4 %(w / v), about 4%(w / v) to about 10%(w / v), about 4%(w / v) to about 9%(w / v), about 4%(w / v) to about 8%(w / v), about 4%(w / v) ) ~ approx. 7% (w / v), approx. 4% (w / v) ~ approx. 6% (w / v), approx. 4% (w / v) ~ approx. 5% (w / v), approx. 5% (w / v) ~ approx. 10% (w / v), approx. 5% (w / v) ~ approx. 9% (w / v), approx. 5% (w / v) ~ approx. 8% (w / v), approx. 5% (w / v) ~ approx. 7% (w / v), approx. 5% (w / v) ~ approx. 6% (w / v) , about 6% (w / v) to about 10% (w / v), about 6% (w / v) to about 9% (w / v), about 6% (w / v) to about 8% (w / v), about 6% (w / v) to about 7% ( and / or one or more blocking agents at a total concentration of about 7% (w / v) to about 10% (w / v), about 7% (w / v) to about 9% (w / v), about 7% (w / v) to about 8% (w / v), about 8% (w / v) to about 10% (w / v), about 8% (w / v) to about 9% (w / v), or about 9% (w / v) to about 10% (w / v).

[0040] In some embodiments, the composition comprises a blocking agent comprising bovine serum albumin at a concentration of about 0.05% (w / v) to about 10% (w / v). In some embodiments, the composition comprises a blocking agent comprising bovine serum albumin at a concentration of about 0.5% (w / v). In some embodiments, the composition comprises a blocking agent comprising bovine serum albumin at a concentration of about 5% (w / v). In some embodiments, the composition comprises a blocking agent comprising bovine serum albumin at a concentration of about 2.5% (w / v).

[0041] In some embodiments, the compositions comprise Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM, calcium chloride (CaCl) at a concentration of about 1 mM to 200 mM, bovine serum albumin (BSA) at a concentration of about 0.05% (w / v) to about 10% (w / v), and one or more block copolymers at a total concentration of about 0.05% (w / v) to 5% (w / v). In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM, sodium chloride (NaCl) at a concentration of 400 mM, calcium chloride (CaCl2) at a concentration of 16 mM, bovine serum albumin (BSA) at a concentration of 5% (w / v), and a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of 0.25% (w / v). In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM, sodium chloride (NaCl) at a concentration of 275 mM, calcium chloride (CaCl2) at a concentration of 8 mM, bovine serum albumin (BSA) at a concentration of 2.5% (w / v), and a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 0.125% (w / v).

[0042] In some embodiments, the composition is present at a concentration of about 0.1% (w / v) to about 3% (w / v), e.g., about 0.1% (w / v) to about 2.5% (w / v), about 0.1% (w / v) to about 2% (w / v), about 0.1% (w / v) to about 1.5% (w / v), about 0.1% (w / v) to about 1% (w / v), about 0.1% (w / v) to about 0.5% (w / v), about 0.1% (w / v) to about 0.4% (w / v), about 0.1% (w / v) to about 0.3% (w / v), about 0.1% (w / v) to about 0.2% (w / v), about 0.2% (w / v) to about 3% (w / v), about 0.2 ... w / v) ~ approx. 2.5% (w / v), approx. 0.2% (w / v) ~ approx. 2% (w / v), approx. 0.2% (w / v) ~ approx. 1.5% (w / v), approx. 0. 2% (w / v) ~ approx. 1% (w / v), approx. 0.2% (w / v) ~ approx. 0.5% (w / v), approx. 0.2% (w / v) ~ approx. 0.4% (w / v), Approx. 0.2% (w / v) ~ approx. 0.3% (w / v), approx. 0.3% (w / v) ~ approx. 3% (w / v), approx. 0.3% (w / v) ~ approx. 2.5% (w / v), about 0.3% (w / v) to about 2% (w / v), about 0.3% (w / v) to about 1.5% (w / v), about 0.3% (w / v) to about 1% (w / v), approx. 0.3% (w / v) ~ approx. 0.5% (w / v), approx. 0.3% (w / v) ~ approx. 0.4% (w / v), approx. 0.4% (w / v) ~ approx. 3% (w / v), about 0.4% (w / v) ~ about 2.5% (w / v), about 0.4% (w / v) ~ about 2% (w / v), about 0.4% (w / v) ~ Approx. 1.5% (w / v), approx. 0.4% (w / v) ~ approx. 1% (w / v), approx. 0.4% (w / v) ~ approx. 0.5% (w / v), approx. 0.5% (w / v) ~ approx. 3% (w / v), approx. 0.5% (w / v) ~ approx. 2.5% (w / v), approx. 0.5% (w / v) ~ approx. 2% (w / v), approx. 0.5% (w The casein concentration is from about 0.5% (w / v) to about 1% (w / v), about 0.5% (w / v) to about 1% (w / v), about 1% (w / v) to about 3% (w / v), about 1% (w / v) to about 2.5% (w / v), about 1% (w / v) to about 2% (w / v), about 1% (w / v) to about 1.5% (w / v), about 1.5% (w / v) to about 3% (w / v), about 1.5% (w / v) to about 2.5% (w / v), about 1.5% (w / v) to about 2% (w / v), about 2% (w / v) to about 3% (w / v), about 2% (w / v) to about 2.5% (w / v), or about 2.5% (w / v) to about 3% (w / v).

[0043] In some embodiments, the composition comprises casein at a concentration of about 0.4% (w / v) to 0.6% (w / v), hi some embodiments, the composition comprises casein at a concentration of 0.5% (w / v).

[0044] In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), sodium chloride (NaCl), calcium chloride (CaCl2), casein, and one or more block copolymers.

[0045] In some embodiments, the compositions comprise Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to 1000 mM, calcium chloride (CaCl) at a concentration of about 1 mM to 200 mM, casein at a concentration of about 0.1% (w / v) to 2% (w / v), and one or more block copolymers at a total concentration of about 0.05% (w / v) to 5% (w / v). In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM, sodium chloride (NaCl) at a concentration of 150 mM, calcium chloride (CaCl2) at a concentration of 16 mM, casein at a concentration of 0.5% (w / v), and a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of 1% (w / v).

[0046] In some embodiments, the block copolymer is a polyoxyethylene-polyoxypropylene block copolymer. In some embodiments, the polyoxyethylene-polyoxypropylene block copolymer is represented by Formula I:

[0047] [ka]

[0048] Formula I wherein x and z=2 to 130, and b=15 to 67. In some embodiments, x and z=120 to 130 and b=50 to 60. In some embodiments, the block copolymer is Synperonic® F108, which has the formula:

[0049] [ka]

[0050] In some embodiments, the composition further comprises one or more chaotropic agents. In some embodiments, the composition further comprises one or more denaturing agents. In some embodiments, the composition further comprises one or more chaotropic agents and denaturing agents.

[0051] In some embodiments, the one or more chaotropic agents and / or denaturing agents are selected from the list consisting of guanidine salts, urea or betaine, hi some embodiments, the one or more chaotropic agents and / or denaturing agents are guanidine hydrochloride.

[0052] In some embodiments, the one or more chaotropic agents and / or denaturing agents are present at a concentration of about 1 mM to about 1000 mM, e.g., about 1 mM to about 900 mM, about 1 mM to about 800 mM, about 1 mM to about 700 mM, about 1 mM to about 600 mM, about 1 mM to about 500 mM, about 1 mM to about 400 mM, about 1 mM to about 300 mM, , about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 10 mM to about 1000 mM, about 10 mM to about 900 mM, about 10 mM to about 800 mM, about 10 mM to about 700 mM, about 10 mM to about 600 mM, about 10 mM to about 500 mM, about 10 mM to about 400 mM, about 10 mM to about 300 mM, about 10 mM to about 200 mM M, about 10mM to about 100mM, about 50mM to about 1000mM, about 50mM to about 900mM, about 50mM to about 800mM, about 50mM to about 700mM, about 5 0mM to about 600mM, about 50mM to about 500mM, about 50mM to about 400mM, about 50mM to about 300mM, about 50mM to about 200mM, about 50mM to about The concentration is 100 mM, about 100 mM to about 1000 mM, about 100 mM to about 900 mM, about 100 mM to about 800 mM, about 100 mM to about 700 mM, about 100 mM to about 600 mM, about 100 mM to about 500 mM, about 100 mM to about 400 mM, about 100 mM to about 300 mM, or about 100 mM to about 200 mM.

[0053] In some embodiments, the compositions comprise Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to 1000 mM, calcium chloride (CaCl2) at a concentration of about 1 mM to 200 mM, casein at a concentration of about 0.1% (w / v) to 2% (w / v), one or more block copolymers at a total concentration of about 0.05% (w / v) to 5% (w / v), and one or more chaotropic agents and / or denaturing agents at a concentration of about 1 mM to about 1000 mM. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 25 mM, sodium chloride (NaCl) at a concentration of 150 mM, calcium chloride (CaCl2) at a concentration of 16 mM, casein at a concentration of 0.5% (w / v), polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of 1% (w / v), and guanidine hydrochloride (guanidine HCl) at a concentration of 200 mM.

[0054] In some embodiments, the compositions comprise Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM, calcium chloride (CaCl) at a concentration of about 1 mM to about 200 mM, bovine serum albumin at a concentration of about 0.5% (w / v) to about 10% (w / v), one or more block copolymers at a total concentration of about 0.05% (w / v) to about 5% (w / v), and one or more chaotropic agents and / or denaturing agents at a concentration of about 1 mM to about 1000 mM. In some embodiments, the composition comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, sodium chloride (NaCl) at a concentration of about 275 mM, calcium chloride (CaCl) at a concentration of about 8 mM, bovine serum albumin at a concentration of about 2.5% (w / v), polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 0.125% (w / v), and guanidine hydrochloride (guanidine HCl) at a concentration of about 300 mM.

[0055] The present invention also provides compositions for immunoassays comprising one or more chaotropic agents and / or denaturing agents. In some embodiments, the one or more chaotropic agents and / or denaturing agents are selected from the list consisting of guanidine salts, urea or betaine. In some embodiments, the one or more chaotropic agents and / or denaturing agents are guanidine hydrochloride.

[0056] In some embodiments, the one or more chaotropic agents and / or denaturing agents are present at a concentration of about 1 mM to about 1000 mM, e.g., about 1 mM to about 900 mM, about 1 mM to about 800 mM, about 1 mM to about 700 mM, about 1 mM to about 600 mM, about 1 mM to about 500 mM, about 1 mM to about 400 mM, about 1 mM to about 300 mM, , about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 10 mM to about 1000 mM, about 10 mM to about 900 mM, about 10 mM to about 800 mM, about 10 mM to about 700 mM, about 10 mM to about 600 mM, about 10 mM to about 500 mM, about 10 mM to about 400 mM, about 10 mM to about 300 mM, about 10 mM to about 200 mM M, about 10mM to about 100mM, about 50mM to about 1000mM, about 50mM to about 900mM, about 50mM to about 800mM, about 50mM to about 700mM, about 5 0mM to about 600mM, about 50mM to about 500mM, about 50mM to about 400mM, about 50mM to about 300mM, about 50mM to about 200mM, about 50mM to about The concentration is 100 mM, about 100 mM to about 1000 mM, about 100 mM to about 900 mM, about 100 mM to about 800 mM, about 100 mM to about 700 mM, about 100 mM to about 600 mM, about 100 mM to about 500 mM, about 100 mM to about 400 mM, about 100 mM to about 300 mM, or about 100 mM to about 200 mM.

[0057] In some embodiments, the composition further comprises a proteinaceous matrix, optionally, the proteinaceous matrix comprises Bovine Serum Albumin (BSA) and / or casein.

[0058] In some embodiments, the composition further comprises a biocide, optionally comprising 2-methyl-4-isothiazolin-3-one (MIT), 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT) or a mixture thereof (e.g., a ProClin™ biocide such as ProClin™ 300).

[0059] In some embodiments, the composition comprises a biocide at a concentration of about 0.01% (w / v) to about 2% (w / v), e.g., about 0.01% (w / v) to about 1% (w / v), about 0.01% (w / v) to about 0.5% (w / v), about 0.01% (w / v) to about 0.1% (w / v), about 0.02% (w / v) to about 0.08% (w / v), about 0.03% (w / v) to about 0.07% (w / v), or about 0.05% (w / v).

[0060] In some embodiments, the composition further comprises an antifoaming agent, optionally comprising a non-silicone antifoaming agent in a polyol-based dispersion (e.g., Antifoam 204).

[0061] In some embodiments, the composition comprises an antifoaming agent at a concentration of about 0.01% (w / v) to about 2% (w / v), e.g., about 0.01% (w / v) to about 1% (w / v), about 0.01% (w / v) to about 0.5% (w / v), about 0.01% (w / v) to about 0.1% (w / v), about 0.02% (w / v) to about 0.08% (w / v), about 0.03% (w / v) to about 0.07% (w / v), or about 0.05% (w / v).

[0062] In some embodiments, the composition further comprises a heterophilic blocking composition (e.g., MAB33 IgG1 / IgG1 Poly), which is commonly used in immunoassays to block heterophilic antibody interference.

[0063] In some embodiments, the composition is a liquid, gel or emulsion. In some embodiments, the composition comprises water. In some embodiments, the composition is lyophilized. A lyophilized composition may comprise a ratio of components suitable for adding a liquid, e.g., water, to obtain the concentrations described herein.

[0064] In some embodiments, the composition is an assay buffer composition. In some embodiments, the composition is an assay diluent composition. In some embodiments, the composition is an assay wash composition. In some embodiments, the assay diluent composition is an analyte capture diluent. In some embodiments, the assay diluent is an analyte detection diluent. The analyte capture diluent can be used in an immunoassay as a diluent during a step that includes contacting the analyte with a capture antibody. The analyte detection diluent can be used in an immunoassay as a diluent during a step that includes contacting the analyte with a detection antibody.

[0065] In some embodiments, the liquid composition comprises the components of the composition in an aqueous solution, suspension, or a combination thereof.Thus, the composition may comprise water with the remaining components dissolved or suspended in water.In some embodiments, the composition may be formed by solubilizing one or more components of the composition provided in lyophilized form.

[0066] The present invention also provides a method for preparing the immunoassay composition described herein, comprising mixing water with an additional component of the composition to obtain the composition. The present invention also provides a method for preparing the immunoassay composition described herein, comprising mixing an aqueous buffer solution with an additional component of the composition to obtain the composition. The additional component may be provided in solid form, as an aqueous solution, or a combination thereof. Prior to mixing with the aqueous buffer solution, one or more additional components may be provided in lyophilized form.

[0067] The invention also provides a method for preparing an immunoassay composition as described herein, the composition comprising a blocking agent, the method comprising obtaining a liquid composition comprising a buffer compound, a salt, one or more block copolymers and optionally one or more additional components of the composition, providing a lyophilized blocking agent, and mixing the liquid composition with the lyophilized blocking agent to obtain the immunoassay composition.

[0068] In some embodiments, where the composition further comprises one or more chaotropic agents and / or denaturing agents, the method further comprises providing the one or more chaotropic agents and / or denaturing agents in lyophilized form, and mixing the liquid composition with the lyophilized blocking agent and the lyophilized one or more chaotropic agents and / or denaturing agents to obtain a composition for immunoassay.

[0069] In the methods for preparing the immunoassay compositions described herein, the components of the compositions may be provided to prepare a composition having the components and / or concentrations of any of the embodiments of the compositions described herein.For example, in some embodiments, the blocking agent is BSA and / or one or more chaotropic agents and / or denaturing agents are guanidine salts, such as guanidine hydrochloride.

[0070] The present invention also provides a method for performing an immunoassay, comprising contacting a biological sample with one or more compositions according to the present invention. In some embodiments of the method, the immunoassay is for detecting the presence of an analyte of interest in the biological sample. In some embodiments of the method, the biological sample is contacted with one or more compositions according to the present invention simultaneously or sequentially.

[0071] In some embodiments of the method, the one or more compositions are used as a diluent for the biological sample.

[0072] In some embodiments of the method, the method includes contacting the biological sample with a capture antibody, and one or more compositions are used as a capture diluent when the biological sample is contacted with the capture antibody. In some embodiments of the method, the method includes adding a detection antibody, and one or more compositions are used as a detection diluent when the detection antibody is added. In some embodiments of the method, the addition of the detection antibody can be performed simultaneously or after the contact of the biological sample with the capture antibody. In some embodiments of the method, a washing step using a washing solution is performed after contacting the biological sample with the capture antibody, and the detection diluent is added to the capture antibody before or simultaneously with the addition of the detection antibody.

[0073] In some embodiments of the method, one or more compositions are used as washing solution.In some embodiments of the method, washing solution comprises or consists essentially of Tris-buffered saline and block copolymer, mutatis mutandis, as defined in any of the embodiments described herein.

[0074] In some embodiments of the method, one or more of the compositions is used as a buffer. In some embodiments of the method, the biological sample is a blood sample, e.g., a whole blood sample or blood fraction, amniotic fluid, aqueous humor, bile, plasma, breast milk, Cerebrospinal Fluid (CSF), endolymph, extracellular fluid, exudate, gastric acid, hemolymph, interstitial fluid, lymph, mucus, pericardial fluid, peritoneal fluid, sweat (perspiration), sputum, pus, saliva, semen, synovial fluid, tears, urine, vaginal fluid, vomit, saliva, other biological fluids, or a swab sample. In some embodiments of the method, the biological sample is a whole blood sample.

[0075] In some embodiments of the method, the biological sample comprises an analyte of interest, and optionally the analyte of interest is a protein.In some embodiments of the method, the analyte of interest is brain natriuretic peptide or N-terminal pro-BNP.In some embodiments of the method, the analyte of interest is cardiac troponin or cardiac troponin subunit I (cTnI).

[0076] In some embodiments of the method, the immunoassay is a competitive sandwich immunoassay. In some embodiments of the method, the immunoassay comprises obtaining an electrochemical measurement to detect an analyte of interest. In some embodiments of the method, the electrochemical measurement is an amperometric, voltametric, potentiometric, impedancemetric, or electrochemical impedance spectroscopy measurement. In some embodiments of the method, the electrochemical measurement is a chronoamperometric measurement.

[0077] In some embodiments of the method, the method is for measuring an analyte of interest in a biological sample, a. A biological sample is a first antibody or antigen-binding portion thereof capable of binding to the analyte of interest; a second antibody or antigen-binding portion thereof capable of binding to the analyte of interest conjugated to an enzyme; and An assay diluent, which is any composition according to the present invention. to form an assay mixture; b. contacting the assay mixture with a substrate for the enzyme, where the enzyme substrate is converted by the enzyme into an electroactive molecule; and c. Obtaining an electrochemical measurement to measure the analyte of interest.

[0078] In some embodiments of the method, the method is for measuring an analyte of interest in a biological sample, a. The biological sample is a magnetically susceptible bead conjugated to a first antibody or an antigen-binding portion thereof capable of binding to the analyte of interest; a second antibody or antigen-binding portion thereof capable of binding to the analyte of interest conjugated to an enzyme; and An assay diluent, which is any composition according to the present invention. to form an assay mixture; b. holding magnetically susceptible beads on electrodes using a magnetic field; c. contacting the magnetically susceptible beads with a substrate of the enzyme, where the enzyme substrate is converted by the enzyme into an electroactive molecule; and d. Obtaining an electrochemical measurement using the electrode.

[0079] In some embodiments of the method, the method is for measuring an analyte of interest in a biological sample, a. mixing the biological sample with magnetically susceptible beads conjugated to a first antibody or antigen-binding portion thereof capable of binding to the analyte of interest in the presence of a capture diluent, the capture diluent being any composition according to the present invention; b. immobilizing magnetically susceptible beads on a magnet and washing the beads; c. providing a detection diluent with a second antibody or antigen-binding portion thereof capable of binding to the analyte of interest conjugated to an enzyme to produce a detection solution, wherein the detection diluent is any composition according to the present invention; d. adding detection solution to the washed magnetically susceptible beads; e. Washing the magnetically susceptible beads; f. contacting the magnetically susceptible bead with a substrate of the enzyme, whereby the enzyme substrate is converted by the enzyme into an electroactive molecule; and g. Obtaining an electrochemical measurement to measure the analyte of interest.

[0080] In some embodiments of the method, the washing of steps b and e is performed with a washing solution, optionally the washing solution being any composition according to the present invention. In some embodiments of the method, the washing solution comprises or consists essentially of Tris-buffered saline and a block copolymer as described herein.

[0081] In some embodiments of the method, the capture diluent is any of the compositions described herein and includes a buffer compound, at least one salt, one or more block copolymers, one or more blocking agents, and one or more chaotropic agents and / or denaturing agents. In some embodiments of the method, the capture diluent includes at least two salts.

[0082] In some embodiments of the method, the capture diluent comprises Tris, NaCl, CaCl2, polyoxyethylene-polyoxypropylene block copolymer (eg, Synperonic® F108), bovine serum albumin and guanidine hydrochloride (guanidine HCl).

[0083] In some embodiments of the method, the detection diluent is any composition according to the invention and includes a buffer compound, at least one salt, one or more block copolymers, and one or more blocking agents. In some embodiments of the method, the detection diluent includes at least two salts. In some embodiments of the method, the detection diluent includes Tris, NaCl, CaCl2, a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108), and bovine serum albumin.

[0084] In some embodiments of the method, one or more compositions according to the invention are prepared by any suitable method described herein.

[0085] In some embodiments of the method, the immunoassay is performed using a microfluidic cartridge, optionally a microfluidic disposable cartridge. In some embodiments of the method, one or more compositions according to the invention are prepared using the microfluidic cartridge by any suitable method described herein. In some embodiments of the method, one or more compositions prepared using the microfluidic cartridge by the methods described herein are capture diluents and / or detection diluents.

[0086] The present invention also provides for the use of compositions according to the present invention in immunoassays. The present invention also provides for the use of compositions according to the present invention in point-of-care diagnostic assays, such as immunoassays. In some embodiments, the immunoassay is a competitive sandwich immunoassay. In some embodiments, the immunoassay comprises obtaining an electrochemical measurement to detect an analyte of interest. In some embodiments, the electrochemical measurement is an amperometric, voltametric, potentiometric, impedancemetric, or electrochemical impedance spectroscopy measurement. In some embodiments, the electrochemical measurement is a chronoamperometric measurement.

[0087] The present invention also provides One or more compositions according to the present invention; Point-of-care immunoassay diagnostic tests and kits comprising the same are provided.

[0088] In some embodiments, the kit further comprises instructional materials, optionally, the instructional materials comprising instructions for carrying out a method of performing an immunoassay according to the present invention.

[0089] Any of the embodiments of the composition, method, use or kit of the present invention may be combined with any other embodiment, mutatis mutandis.For example, the concentration described for each of the various components of the composition may be combined with any of the concentrations for any of the other components of the concentration.This disclosure describes the composition of the present invention suitable for use in immunoassay.This disclosure also represents the disclosure of any composition itself described herein.

[0090] Embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which: [Brief description of the drawings]

[0091] [Figure 1]Comparison of chronoamperometric measurements of human whole blood samples obtained with the multiwell plate device. Some surfactants were formulated differently and the identity of the surfactant is noted in the figure legend. [Diagram 2] Comparison of chronoamperometric measurements of human whole blood samples obtained with microfluidic disposable cartridges. Some surfactants were formulated differently and the identity of the surfactant is noted in the figure legend. [Diagram 3] Calibration of immunoassay response to analyte in human whole blood samples. Two surfactants, diluent buffer and wash buffer, differ in formulation and are indicated in the figure legend as to which surfactant they are. Dilution buffer was spiked with various concentrations of analyte, ranging from 0 to 200 pg / mL. For each spiked concentration, the current signal was estimated for 45 s. Linear fitting to the concentration response is presented for the two formulations. [Figure 4] Sensitivity of immunoassay response to analytes in human whole blood samples. Two detergents, diluent buffer and wash buffer, differed in formulation and are indicated in the figure legend as to which detergent they were. Dilution buffer was spiked with various concentrations of analyte, ranging from 0 to 200 pg / mL. For each time point in the chronoamperometric trace, a linear model was used to estimate the slope of the concentration response. In all cases, the Synperonic® formulation showed greater sensitivity compared to the Tween formulation. [Diagram 5] Background of immunoassay response to analyte in human whole blood samples. Two detergents, diluent buffer and wash buffer, differ in formulation and are identified in the figure legend. Dilution buffer was spiked with various concentrations of analyte, ranging from 0 to 200 pg / mL. For each time point in the chronoamperometric trace, a linear model was used to estimate the intercept of the concentration response. While the Synperonic® formulation shows a relatively flat background signal, for the Tween formulation the background rises quite sharply after 20 seconds. [Figure 6]Synperonic® titration in dilution buffer containing Bovine Serum Albumin (BSA) showing increased assay sensitivity with increasing Synperonic® concentration. Increasing the Synperonic® concentration in the sample diluent increases the difference in assay response (chronoamperometric current after 60 seconds) between samples containing no analyte and samples containing 200 ng / L analyte. [Figure 7] Synperonic® titration in diluent buffer containing casein (Bovine Serum Albumin (BSA)) showing increased assay sensitivity with increasing Synperonic® block copolymer concentration. Increasing the Synperonic® concentration in the sample diluent increases the difference in assay response (chronoamperometric current after 60 seconds) between samples containing no analyte and samples containing 200 ng / L analyte. [Figure 8] Calibration of immunoassay responses to analytes using dilution buffer containing Bovine Serum Albumin (BSA) and Synperonic® block copolymers in 96-well plates. [Figure 9] Calibration of immunoassay response to analyte using a diluent buffer containing Bovine Serum Albumin (BSA) and Synperonic® block copolymer in a microfluidic disposable cartridge. [Figure 10] Calibration of immunoassay responses to analytes using diluent buffers containing casein and Synperonic® block copolymers in microfluidic disposable cartridges [Figure 11] Calibration curve for cardiac troponin (cTnI) assay performed in 96-well plates using diluents formed from liquid and lyophilized components DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0092] The methods described above may be performed in combination with one another, in a variety of different orders, or multiple times, as necessary for a given diagnostic test, and one method need not be completed before another method is performed.

[0093] Block Copolymers A block copolymer contains two or more homopolymer subunits linked by covalent bonds. The linking of the homopolymer subunits may require an intermediate non-repeating subunit known as a linking block. Diblock copolymers have two distinct blocks and triblock copolymers have three. A block is a portion of a macromolecule that contains many units that have at least one characteristic that is not present in the adjacent portion. One possible sequence of repeat units A and B in a triblock copolymer can be ~AAAAAAABBBBBBBAAAAA~. In some embodiments of the present invention, the block copolymer may have one or more different blocks, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or more different blocks. In a preferred embodiment, the block copolymer is a triblock copolymer comprising three different blocks. The number of repeating monomer units in each block may be any number that allows the block copolymer to retain at least one desired property. In some embodiments of the invention, the number of repeating monomer units in each block is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, It can be 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 200, 300, 400, 500, 1000, 2000, 3000, 4000, 5000, 10000 or more.

[0094] Block copolymers are composed of blocks of different polymerized monomers. For example, polystyrene-b-poly(methyl methacrylate) or PS-b-PMMA (b=block) is usually produced by first polymerizing styrene and then methyl methacrylate (MMA) from the reactive end of the polystyrene chain. Triblocks, tetrablocks, multiblocks, etc. can also be produced. Diblock copolymers are produced using living polymerization techniques such as Atom Transfer free Radical Polymerization (ATRP), Reversible Addition Fragmentation chain Transfer (RAFT), Ring-Opening Metathesis Polymerization (ROMP), Chain Shuttle Polymerization and Living Cationic or Living Anionic Polymerization.

[0095] The "blockiness" of a copolymer is a measure of the proximity of the comonomers relative to the statistical distribution of the comonomers. Many, and indeed most, synthetic polymers are in fact copolymers containing a minority of monomers, about 1-20%. In such cases, blockiness is undesirable. A quantitative measure of blockiness or deviation from random monomer composition has been proposed: block index.

[0096] In some embodiments, the block copolymers used in the present invention are poloxamers (also known as Pluronics® or Synperonics®). Poloxamers are a class of synthetic block copolymers consisting of hydrophilic poly(ethylene oxide) (PEO) and hydrophobic poly(propylene oxide) (PPO) arranged in an ABA triblock structure, thus giving PEO-PPO-PEO. They can be obtained in either liquid, paste or solid form. Due to their amphiphilic character (the presence of hydrophobic and hydrophilic components), poloxamers have surfactant properties that allow them to interact with hydrophobic surfaces and biological molecules. Being amphiphilic also results in the ability of the individual block copolymers, known as monomers, to combine in aqueous solution to form micelles. When the concentration of the block copolymer is below the critical micelle concentration (CMC), the monomers remain in molecular solution in water. However, when the block copolymer concentration is increased above the CMC, the monomers self-assemble to form micelles, which can adopt spherical, rod-like or lamellar shapes. Their shape depends on the length and concentration of the block copolymers (i.e., EO, ethylene oxide and PO, propylene oxide) and the temperature. Micelles usually have a hydrophobic core, in this case PO chains, and a hydrophilic shell, EO chains. [1] Poloxamers are represented by formula I:

[0097] [ka]

[0098] Formula I where x, y and z are integer values, preferably x and z=2-130 and b=15-67. In some embodiments, x and z may be preferably 120-130 and b may be preferably 50-60. Poloxamers are generally named by the letter P (for poloxamer) followed by three digits, where the first two digits multiplied by 100 indicate the approximate molecular mass of the polyoxypropylene core and the last digit multiplied by 10 indicates the polyoxyethylene content (e.g., P407=a poloxamer with a polyoxypropylene molecular mass of 4000 g / mol and a polyoxyethylene content of 70%). The average molecular weight and weight percentage of polyoxyethylene of the poloxamers can be determined according to the United States Pharmacopeia (United States Pharmacopeia (2022), NF Monographs, Poloxamers, USP-NF, Rockville, MD; DOI: https: / / doi.org / 10.31003 / USPNF_M66210_05_01, incorporated herein by reference). In the case of Pluronics® and Synperonics®, the nomenclature of these copolymers begins with a letter that defines its physical form at room temperature (L=liquid, P=paste, F=flake (solid)), followed by two or three digits, where the first digit of the numerical designation (two digits for three-digit numbers) multiplied by 300 indicates the approximate molecular weight of the hydrophobe, and the last digit x 10 indicates the polyoxyethylene content.

[0099] In some embodiments, the block copolymer has a formula according to Formula I.

[0100] [ka]

[0101] Formula I And Formula I is poloxamer P338 (eg, Synperonic® F108).

[0102] Poloxamer P338 may be as defined in the United States Pharmacopoeia, i.e. having an average molecular weight of 12700 to 17400 and an oxyethylene content of 83.1±1.7% by weight. Synperonic® F108 denotes a copolymer that is solid at room temperature (F-flake) and has a molecular mass of polyoxypropylene of 3000 g / mol, corresponding to a polyoxyethylene content of 80%.

[0103] Chaotropic or Denaturing Agents Chaotropic agents are substances that disrupt and denature the structure of macromolecules such as proteins and nucleic acids (e.g., DNA and RNA). Chaotropic solutes increase the entropy of a system by interfering with intermolecular interactions mediated by noncovalent forces such as hydrogen bonding, van der Waals forces, and the hydrophobic effect.

[0104] The structure and function of macromolecules depend on the net effect of these forces, therefore an increase in chaotropic solutes in biological systems will denature macromolecules, reduce enzyme activity, and induce stress on cells.

[0105] Tertiary protein folding depends on the hydrophobicity of amino acids throughout the protein's sequence. Chaotropic solutes reduce the net hydrophobic effect of hydrophobic regions due to disordering of water molecules adjacent to the protein. This solubilizes the hydrophobic regions in solution, thereby denaturing the protein. This also applies to hydrophobic regions of lipid bilayers, and when a critical concentration of chaotropic solutes is reached (in the hydrophobic regions of the bilayer), membrane integrity is compromised and cells are lysed.

[0106] Chaotropic salts that dissociate in solution exert their chaotropic effect through different mechanisms. While chaotropic compounds such as ethanol interfere with non-covalent intramolecular forces as outlined above, salts can have chaotropic properties by shielding charges and preventing the stabilization of salt bridges. Since hydrogen bonds are stronger in non-polar media, salts that increase the chemical polarity of the solvent can also destabilize hydrogen bonds. Mechanistically, this is because there are insufficient water molecules to effectively solvate the ions. This can result in ion-dipole interactions between the salt and hydrogen-bonded species that are more favorable than regular hydrogen bonds.

[0107] Examples of chaotropic or denaturing agents suitable for use in the compositions of the invention include guanidine salts (such as guanidine hydrochloride), betaine, urea, n-butanol, ethanol, guanidinium chloride, lithium perchlorate, lithium acetate, magnesium chloride, phenol, 2-propanol, sodium dodecyl sulfate, and thiourea.

[0108] Biological samples In the present invention, the sample liquid can be any suitable biological sample that contains the diagnostic biomarker of interest. In some embodiments, the sample liquid can be a whole blood sample, a serum sample, a saliva sample, a biopsy sample (such as a healthy tissue sample or a tumor sample), a urine sample, a semen sample, a tear sample, a sputum sample, a sweat sample, a mucous membrane sample, a fecal sample, a gastric fluid sample, an abdominal fluid sample, an amniotic fluid sample, a cyst fluid sample, a peritoneal fluid sample, a spinal fluid sample or a synovial fluid sample, although a whole blood sample is particularly useful. In a preferred embodiment of the present invention, the sample liquid is a whole blood sample. The method may include a step of obtaining or preparing a biological sample, or the sample can be previously obtained from a subject, for example, by an ex vivo method.

[0109] Biological samples obtained from subjects can be stored until needed. Suitable storage methods include freezing immediately after sample collection within 2 hours or up to 2 weeks. Maintenance at -80°C can be used for long-term storage. Preservatives can be added or samples can be collected in tubes containing preservatives. Preferably, samples are analyzed immediately after collection.

[0110] The method of the present invention may include a step performed on a biological sample. The sample liquid is considered to represent the biomarker status of the biomarker of interest in different disease states. Thus, the method of the present invention can use quantitative data on the biomarker of interest to determine the presence or absence or severity of various disease states.

[0111] The sample can be processed before determining the status of the biomarker. The sample can be subjected to concentration (e.g., to increase the concentration of the biomarker to be quantified), centrifugation or dilution. In other embodiments, the sample does not undergo pretreatment and is used raw (e.g., whole blood).

[0112] In some embodiments of the present invention, biological samples may be fractionated or enriched for specific biomarkers prior to detection and quantification (i.e., measurement). The fractionation or enrichment step may be any suitable pretreatment method step to increase the concentration of the biomarker of interest in the sample. For example, the fractionation and / or enrichment step may include centrifugation and / or filtration to remove cells or undesired analytes from the sample, or to increase the concentration of the biomarker of interest in a particular blood fraction. Such methods may be used to enrich a sample for any biomarker of interest.

[0113] The method of the present invention can be carried out on one test sample from a subject. Alternatively, multiple test samples can be taken from a subject, for example, at least two, at least three, at least four or at least five samples can be taken from a subject. Each sample can be subjected to a single assay to quantify one of the biomarker panel members, or the sample can be tested for all of the biomarkers to be quantified. Each sample can be subjected to separate analysis using the method of the present invention, or the method can include multiple samples from a single subject undergoing diagnosis.

[0114] "Sample(s)", "sample(s)", sample liquid, or "sample(s) of interest" are terms used interchangeably in the singular or plural, are not intended to be limited to a particular amount, and as used herein may be any molecule or substance about which a user wishes to gather information. A sample may become larger or smaller in size, volume, or content during the performance of an assay (e.g., by expansion or division, respectively). Thus, a sample may be amplified and / or fractionated one or more times during the performance of an assay. In some embodiments, the sample contains a biomarker of interest.

[0115] As used herein, "liquid" is any aqueous or lipophilic phase that can flow freely.Liquid may further comprise one or more compositions of interest, reaction components or samples selected from cells (including, but not limited to, any eukaryotic or prokaryotic cells, such as cells selected from human, animal, plant, fungus, bacteria, virus, protozoa, yeast, mold, algae, rickettsia and prion), proteins, peptides, antibodies, nucleic acid sequences, oligonucleotide probes, polymerase enzymes, buffers, dNTPs, organic and inorganic chemicals, and fluorescent dyes.

[0116] Assay Method Diagnostic immunoassays suitable for use with the compositions of the present invention can be implemented by diagnostic systems using computer executable instructions. A computer program product or computer readable medium can include or store computer executable instructions. A computer program product or computer readable medium can include hard disk drives, flash memory, read-only memory (ROM), CDs, DVDs, cache, random access memory (RAM), and / or any other storage medium in which information is stored for any period of time (e.g., for a long period of time, permanently, for a short period of time, as temporary buffering, and / or as information cache). A computer program can include computer executable instructions. A computer readable medium can be a tangible or non-transitory computer readable medium. The term "computer readable" encompasses "machine readable".

[0117] Thus, also disclosed is a computer program comprising computer executable instructions that, when executed by a diagnostic system, causes the diagnostic system to perform a diagnostic immunoassay suitable for use with the compositions of the present invention.

[0118] The present embodiments are not limited to the microfluidic scale and other, e.g., macroscopic scale applications are envisaged as well. For the avoidance of doubt, the term "microfluidic" is intended herein to mean a device having fluidic elements such as reservoirs or channels having at least one dimension less than 1 mm.

[0119] Example 1 - Use of block copolymer compositions in stabilizing electrochemical signals Many biosensors that operate on blood use immunoassays, often including sandwich ELISAs, as their detection mechanism. The end of the sandwich can be a detection enzyme (e.g., a peroxidase enzyme such as horseradish peroxidase-HRP), which is the component that ultimately generates the sensor's signal.

[0120] However, blood components contain hemoglobin, as well as other enzymes such as myeloperoxidase, NADPH oxidase, etc., that may be present at the moment of signal generation. These may act as peroxidase mimics, so their presence may interfere with the measurement and lead to erroneous results. Their presence also reduces the quality of the electrochemical signal, from a smooth and clear response to a very irregular and noisy one. This is believed to be because the series of incubation steps, washing steps and exchanges of solutions leads to the lysis of blood cells remaining on the assay or sensor, thereby releasing the enzymes mentioned above.

[0121] The inventors have discovered that by manipulating the type of surfactant and the concentration of the surfactant in the diluent and wash solutions, it is possible to alter the role that hemoglobin plays in electrochemical measurements.

[0122] The typical approach in the art is to use Tween-20 as diluent and washing solution, which results in unstable and irregular electrochemical traces. By modifying the diluent to contain block copolymers (such as Synperonic® F108 or pluronics), the onset of enzyme interference in the signal can be delayed, allowing easier measurement and processing of the signal trace, and thus achieving more accurate diagnosis.

[0123] The use of block copolymers has significant advantages for the following reasons: (i) block copolymers are fully synthetic and highly stable components in aqueous solution, (ii) the electrochemical signal is cleaner, smoother and easier to process, reducing overall measurement-related errors, (iii) it is possible to measure the electrochemical signal over a longer time frame which may result in higher assay sensitivity, and (iv) these block copolymers can be introduced into the wash buffer and also act as a strong detergent to remove unbound analyte and reduce non-specific signal from the electrode.

[0124] Use of the compositions according to the present invention can also reduce the background signal seen in electrochemical detection.

[0125] Example 2 - Dilution Buffer Containing Block Copolymer as Emulsifier In most assays, the formulation plays a very important role in the performance of the assay. Most standard assays use a diluent that contains one or more of the following compositions: a buffer, a protein source, a detergent, and additional components to reduce non-specific interactions. The ratio and selection of components is important because they affect many aspects of the assay, such as antibody kinetics, sensitivity, precision and reproducibility, and help reduce sample-to-sample variability.

[0126] Assays should be developed to have fast binding kinetics suitable for point-of-care (PoC) applications, as well as small donor-to-donor variability. Furthermore, for the assay to work with a wide range of samples (e.g., serum, plasma, and whole blood), it is necessary that the formulation works effectively regardless of the sample matrix and provides consistent and comparable results.

[0127] One such composition includes casein. Casein is widely used as an immunoassay blocker due to its protein bulking effect as well as its low cost. It is a hydrophobic protein that can generate micelles in aqueous solution under certain conditions. The problem is that proteins from the matrix as well as antibodies and enzymes, especially those with large molecular weights, can easily aggregate in such solutions.

[0128] A preferred method that can be implemented using the compositions of the present invention uses enzymes linked to high molecular weight polymers to amplify electrochemical signals. Furthermore, these polymer-enzyme components are used at high concentrations to improve assay kinetics and enhance sensitivity. However, these components can generally aggregate in solution, especially when whole blood is used as the matrix.

[0129] The solution to this problem is the use of block copolymers, which act as emulsifiers. Block copolymers are synthetic molecules that usually contain two hydrophilic and one hydrophobic part. These components can be tailored with monomer blocks of different lengths to give different solubilities in aqueous and hydrophobic environments, usually measured by the HLB index hydrophilic-lipophilic balance.

[0130] The use of PEO-PPO-PEO block copolymers to create the emulsion allows for better solubilization of the whole blood matrix as well as the polymer-enzyme components, which has a significant impact on increasing the sensitivity of the assay as well as reducing donor-to-donor variability.

[0131] Additional advantages include the stability of these compositions at room temperature. Moreover, by varying the molecular weight and HLB of the polymer, it is possible to modify the composition to function as multiple biomarkers and in various assays. Furthermore, since these polymers act as detergents, there is no need to add additional detergents to the formulation. Finally, it also acts as a wetting agent.

[0132] The combination of well-known materials for immunoassays, such as casein, with polymeric materials to create emulsions to improve assay performance has not been disclosed in the prior art, especially for assays utilizing large polymeric recognition molecules for highly sensitive assay applications.

[0133] Example 3 - Dilution buffer containing a guanidine salt Non-specific binding is a common problem in immunoassays. This may manifest as a higher than expected signal, which not only affects the sensitivity and limit of detection (LoD) of the immunoassay, but also leads to erroneous results from samples of unknown concentration. A preferred assay that can be performed using the composition of the present invention uses a high molecular weight polymeric enzyme component to increase sensitivity, and in some cases, the background electrochemical signal of the assay may be significantly elevated. In addition, clinical samples may also aggregate with antibodies due to the presence of proteins that non-specifically bind to the antibody-antigen complex. This also manifests as a higher background non-specific signal that causes assay inaccuracy.

[0134] The use of low concentrations of strong denaturants and chaotropes, such as guanidine hydrochloride, urea and betaine, traditionally used in molecular biology to denature nucleic acid templates, can also have a beneficial effect in reducing background signal in immunoassays.

[0135] This background reduction is also sometimes accompanied by a reduction in the specific signal, but background reduction is much more preferable. For example, guanidine salts are particularly effective in reducing the background of cardiac troponin I assays for at least the following reasons: (i) considering the requirements of high sensitivity and low detection limits, guanidine salts disrupt complexes that cause non-specific signals, (ii) significantly reduce aggregation of large enzyme polymers that may form especially at low analyte concentrations, and (iii) slightly denature the troponin antigen, thereby resulting in better accessibility of antibodies and reducing the effect of other troponin-binding proteins, which may in turn result in non-specific interactions together. Guanidine salts and similar chaotropes work best when added to a protein matrix (BSA, casein).

[0136] Other advantages of such compositions include their exceptional stability in a variety of conditions, making them suitable components for point-of-care (PoC) devices where shelf life in a variety of environmental conditions is critical.

[0137] Example 4 - Wash Buffer with Block Copolymer Figure 1 shows the effect on the quality of the electrochemical signal by adding different surfactants to the wash buffer used to perform the immunoassay on a 96-well plate. The sample was human whole blood mixed with diluent (25 mL Tris buffer, 400 mM NaCl, 16 mM CaCl2, 200 mM guanidine hydrochloride, 1% casein, 0.03% heterophilic blocking composition-24), 4ug / mL of detection antibody and magnetic beads coated with capture antibody. This mixture was incubated on the 96-well plate at 37°C for several minutes. The mixture was then emptied and washed twice with 100uL, retaining the magnetic beads each time. The wash buffer was Tris-Buffered Saline (TBS) mixed with 0.1% surfactant, the identity of which is indicated in the figure legend (see Figure 1).

[0138] After washing, the magnetic particles were localized on a 3 mm electrode and 3,3',5,5'-tetramethylbenzidine (TMB) solution was introduced. Chronoamperometric measurements were performed using a three-electrode system with screen-printed carbon for each of the working, counter and reference electrodes. A voltage of -200 mV was applied for 60 seconds and the current was recorded.

[0139] [Table 1]

[0140] Table 1 - Surfactants investigated. Non-ionic surfactants selected from the Surf's Up® kit from QED Bioscience Inc. Example 5 - Wash Buffer with Block Copolymer Figure 2 shows the effect on the quality of the electrochemical signal by adding different surfactants to the wash buffer used to perform the immunoassay on the microfluidic disposable cartridge. The sample was human whole blood mixed with dilution buffer (25 mL Tris buffer, 400 mM NaCl, 16 mM CaCl2, 200 mM guanidine hydrochloride, 1% casein, 0.03% heterophilic blocking composition-24), 4ug / mL of detection and capture antibody coated magnetic beads. This mixture was incubated in the cartridge for a few minutes while the fluid was manipulated using a home-built device. The magnetic beads were pulled down onto a magnet and washed with 200uL of Tris-Buffered Saline (TBS) + 0.05% of the different surfactants as indicated in the figure legend. After washing, the beads were transferred to the second sensor using TBS-0.05% surfactant. The magnetic beads were held in place using a second magnet. Chronoamperometric measurements were performed using a three-electrode system with screen-printed carbon for each of the working, counter and reference electrodes. A voltage of -50 mV was applied for 60 seconds and the current was recorded (see Figure 2).

[0141] [Table 2]

[0142] Table 2 - Surfactants investigated. Non-ionic surfactants selected from the Surf's Up® kit from QED Bioscience Inc. Example 6 - Wash Buffer Containing Block Copolymer Figures 3-5 show the effect of adding different surfactants to the wash buffer on the performance of the immunoassay. The samples were human whole blood mixed with diluent (25 mL Tris buffer, 400 mM NaCl, 16 mM CaCl2, 200 mM guanidine hydrochloride, 1% casein, 0.03% heterophile blocking composition-24), 4 ug / mL detection antibody, 8 ug / mL horseradish peroxidase (HRP) and magnetic beads coated with capture antibody. Various concentrations of analyte, from 0 to 200 pg / mL, were added to the dilution buffer. These mixtures were incubated on a 96-well plate at 37°C for several minutes. The wells were then emptied of the mixtures and washed twice with 100 uL of wash solution, retaining the magnetic beads each time. The washing solution was Tris-Buffered Saline (TBS) mixed with 0.05% detergent, the identity of the detergent being indicated in the figure legends (see Figures 3-5).

[0143] After washing, the magnetic particles were localized on a 3 mm electrode and 3,3',5,5'-tetramethylbenzidine (TMB) solution was introduced. Chronoamperometric measurements were performed using a three-electrode system with screen-printed carbon for each of the working, counter and reference electrodes. A voltage of -200 mV was applied for 60 seconds and the current was recorded.

[0144] [Table 3]

[0145] Table 3 - Surfactants investigated. Non-ionic surfactants selected from the Surf's Up® kit from QED Bioscience Inc. Example 7 - Dilution Buffer with Blocking Agent and Block Copolymer Figure 6 shows the effect on assay performance of increasing concentrations of Synperonic® in a diluent buffer containing Bovine Serum Albumin (BSA). Analyte-free human plasma (lithium heparin) and the same plasma supplemented with 200 ng / L of analyte were measured in parallel using the standard procedure in a 96-well plate format, varying only the concentration of Synperonic® block copolymer in the sample diluent.

[0146] Various sample diluents (aqueous) containing identical concentrations of bovine serum albumin (BSA), sodium chloride (NaCl), calcium chloride (CaCl2), and various concentrations of Synperonic® block copolymer were tested under standardized assay conditions. Plasma samples were mixed with sample diluents containing detection antibodies labeled with horseradish peroxidase (HRP) in a 1:1 ratio and magnetic particles coated with capture antibodies. The reaction mixture was incubated in a well plate at 37°C for 8 minutes, after which the magnetic particles were removed by a magnetic field and washed with an assay wash buffer containing a standard concentration of Synperonic® in water. The washed magnetic particles were localized on a 3 mm electrode and a 3,3',5,5'-tetramethylbenzidine (TMB) solution was introduced. The assay response currents recorded using chronoamperometry at -200 mV for analyte-free and 200 ng / L analyte samples using each sample diluent (varying concentrations of Synperonic®) showed that increasing the concentration of Synperonic® in the sample diluent improved the assay sensitivity by increasing the difference in assay response between analyte-free and 200 ng / L analyte-containing samples (Figure 6).

[0147] Example 8 - Dilution Buffer with Blocking Agent and Block Copolymer Figure 7 shows the effect on assay performance of increasing concentrations of Synperonic® in the diluent buffer containing casein. Analyte-free human plasma (lithium heparin) and the same plasma supplemented with 200 ng / L of analyte were measured in parallel using the standard procedure in a 96-well plate format, varying only the concentration of Synperonic® block copolymer in the sample diluent.

[0148] Various sample diluents (aqueous) containing identical concentrations of casein, sodium chloride (NaCl), calcium chloride (CaCl2), and various concentrations of Synperonic® block copolymer were tested under standardized assay conditions. Plasma samples were mixed with sample diluents containing horseradish peroxidase (HRP)-labeled detection antibodies in a 1:1 ratio and magnetic particles coated with capture antibodies. The reaction mixture was incubated in a well plate at 37°C for 8 minutes, after which the magnetic particles were removed by a magnetic field and washed with an assay wash buffer containing a standard concentration of Synperonic® in water. The washed magnetic particles were localized on a 3 mm electrode and a 3,3',5,5'-tetramethylbenzidine (TMB) solution was introduced. The assay response currents recorded using chronoamperometry at -200 mV for analyte-free and 200 ng / L analyte samples using each sample diluent (varying concentrations of Synperonic®) showed that increasing the Synperonic® concentration in the sample diluent improved the assay sensitivity by increasing the difference in assay response between analyte-free and 200 ng / L analyte-containing samples (Figure 7).

[0149] Example 9 - Effect of adding chaotropic agents to the diluent buffer on background from different donors Analyte-free human blood was measured using standard procedures in a 96-well plate format, varying only the concentration of guanidine HCl in the sample diluent.

[0150] The variability of sample diluent (25 mM Tris in water) containing the same concentration of casein (1%), 150 mM sodium chloride (NaCl), 16 mM calcium chloride (CaCl2) and two concentrations of guanidine HCl was investigated under standardized assay conditions (0 mM and 100 mM). Plasma samples were mixed with sample diluent containing detection antibody labeled with horseradish peroxidase (HRP) in a 1:1 ratio and magnetic particles coated with capture antibody. The reaction mixture was incubated in a well plate at 37°C for 8 minutes, after which the magnetic particles were removed by a magnetic field and washed with a washing buffer containing 0.05% (w / v) Synperonic®. The washed magnetic particles were localized on a 3 mm electrode and a 3,3',5,5'-tetramethylbenzidine (TMB) solution was introduced. The assay response current recorded at 60 s using chronoamperometry at -200 mV with each sample diluent (varying guanidine HCl) showed that increasing the guanidine HCl concentration in the sample diluent improved the assay sensitivity by decreasing the signal background (Table 2).

[0151] [Table 4]

[0152] Table 4 - Effect of adding guanidine hydrochloride to the diluent buffer on background from different donors Example 10 - Calibration of assay response to analyte using a diluent containing a blocking agent and a block copolymer.

[0153] Figure 8 shows the calibration curve of the immunoassay in a 96-well plate. Analyte-free human plasma (EDTA) and the same plasma supplemented with various concentrations of analyte were measured using standard procedures.

[0154] The magnetic particles coated with the capture antibody were mixed with the plasma sample, sample diluent (25 mM aqueous TBS) containing bovine serum albumin (5%), sodium chloride (400 mM), calcium chloride (16 mM) and Synperonic® block copolymer (0.25%), and detection antibody labeled with horseradish peroxidase (HRP). The reaction mixture was incubated in the well plate at 37° C. for 5 min, after which the magnetic particles were removed by a magnetic field and washed with an assay wash buffer containing a standard concentration of Synperonic® in water. The washed magnetic particles were localized on an electrode and a 3,3′,5,5′-tetramethylbenzidine (TMB) solution was introduced. The assay response recorded for each sample (current chronoamperometry after 60 s at −200 mV) was plotted against the analyte concentration to estimate the sensitivity of the assay response to the analyte (FIG. 8).

[0155] Example 11 - Calibration of assay response to analyte using a diluent containing a blocking agent and a block copolymer.

[0156] Figure 9 shows the calibration curve of the immunoassay in the microfluidic disposable cartridge. Analyte-free human plasma (EDTA) and the same plasma supplemented with various concentrations of analyte were measured using standard procedures.

[0157] The magnetic particles coated with the capture antibody were mixed with the plasma sample, sample diluent (25 mM aqueous TBS) containing bovine serum albumin (5%), sodium chloride (400 mM), calcium chloride (16 mM) and Synperonic® block copolymer (0.25%), and detection antibody labeled with horseradish peroxidase (HRP). The reaction mixture was incubated in the flow channel at 37° C. for 5 min, after which the magnetic particles were removed by a magnetic field and washed with an assay wash buffer containing a standard concentration of Synperonic® in water. The washed magnetic particles were localized on an electrode and a 3,3′,5,5′-tetramethylbenzidine (TMB) solution was introduced. The assay response recorded for each sample (current chronoamperometry after 60 s at −200 mV) was plotted against the analyte concentration to estimate the sensitivity of the assay response to the analyte (FIG. 9).

[0158] Example 12 - Calibration of assay response to analyte using a diluent containing a blocking agent and a block copolymer.

[0159] Figure 10 shows the calibration curve of the immunoassay in the microfluidic disposable cartridge. Analyte-free human plasma (EDTA) and the same plasma supplemented with various concentrations of analyte were measured using standard procedures.

[0160] The magnetic particles coated with the capture antibody were mixed with the plasma sample, sample diluent (25 mM aqueous TBS) containing casein (0.5%), sodium chloride (150 mM), calcium chloride (16 mM) and Synperonic® block copolymer (1%), and detection antibody labeled with horseradish peroxidase (HRP). The reaction mixture was incubated in the flow channel at 37° C. for 5 min, after which the magnetic particles were removed by a magnetic field and washed with an assay wash buffer containing a standard concentration of Synperonic® in water. The washed magnetic particles were localized on an electrode and a 3,3′,5,5′-tetramethylbenzidine (TMB) solution was introduced. The assay response recorded for each sample (current chronoamperometry after 60 s at −200 mV) was plotted against the analyte concentration to estimate the sensitivity of the assay response to the analyte.

[0161] Example 13 - Generation of a calibration curve for cardiac troponin (cTnI) assay using a diluent formed from lyophilized components FIG. 11 shows a calibration curve performed in a 96-well plate using samples of known hs-cTnI concentrations as follows:

[0162] This assay utilized lyophilized beads of reagents. Lyophilized bead 1 contains BSA and magnetic particles coated with capture antibody. Lyophilized bead 2 contains BSA and detection antibody (anti-cTnI conjugated with HRP). Lyophilized bead 3 contains BSA, heterophilic blocking composition and guanidine HCl.

[0163] Lyophilized beads 1 and 3 (of the composition shown in Table 5) were dissolved in 25 μL of assay diluent to produce magnetic particles coated with capture antibodies in analyte capture diluent. Analyte capture diluent containing magnetic particles was used to dissolve 50 μL of sample to generate a uniform reaction volume of 75 μL. After 5 minutes, the magnetic beads were immobilized using a magnet and washed with 400 μL of wash buffer.

[0164] 50 μL of assay diluent (of the composition shown in Table 5) was used to dissolve the lyophilized beads 2 to generate a detection solution containing anti-cTnI detection antibody conjugated with an enzyme label in analyte detection diluent. This detection solution was added to the washed magnetic particles for 2.5 minutes, followed by a second wash step of 400 μL wash buffer. Finally, 250 μL of TMB solution was added on top of the magnetic particles to generate an enzyme product that can be detected by an electrode and is proportional to the analyte concentration. The assay response recorded for each sample (current chronoamperometry after 60 seconds at -200 mV) is plotted against the analyte concentration to estimate the sensitivity of the assay response to the analyte.

[0165] [Table 5]

[0166] Table 5 - Assay diluent reagents and concentrations. Diluents were prepared using Pierce™ 20X Tris-buffered saline (TBS, from Thermo Fisher), sodium chloride, calcium chloride, Synperonic® F108, Antifoam 204, Proclin™ 300 bovine serum albumin and guanidine hydrochloride (from Sigma-Aldrich) and MAB33 IgG1 / IgG1 Poly (from Roche). In the assay diluent, 150 mM NaCl was obtained from TBS and 125 mM NaCl was added.

[0167] A cTnI assay as described above can be performed using a microfluidic cartridge, where the assay reagents may be stored on the cartridge either as liquids (eg, in liquid blisters) or as dry lyophilized beads.

[0168] The embodiments of the present invention described herein and shown in the drawings are merely exemplary embodiments and are not intended to limit the scope of the appended claims, including any equivalents thereto. Various modifications are possible and will be readily apparent to those skilled in the art. Any combination of non-mutually exclusive features described herein is intended to be within the scope of the present invention. That is, features of the described embodiments can be combined with any suitable aspect described above, and any feature of any one aspect can be combined with any other suitable aspect.

[0169] Further embodiments of the present invention are described below. 1. A composition for an immunoassay comprising a buffer compound, a salt, and one or more block copolymers.

[0170] 2. The buffer compounds are Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), Bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), Tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid), HEPES (4 2. The composition for use in an immunoassay according to embodiment 1, wherein the carboxylate is selected from the list consisting of N,N'-bis(2-ethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), cacodylic acid (dimethylarsenic acid) and MES (2-(N-morpholino)ethanesulfonic acid).

[0171] 3. Salts include sodium chloride, aluminum ammonium sulfate, aluminum chloride, aluminum chloride, aluminum fluoride, aluminum nitrate, aluminum potassium sulfate, aluminum sulfate, antimony(III) sulfide, antimony trichloride, barium bromide, barium carbonate, barium chloride, barium chromate, barium nitrate, barium perchlorate, barium sulfate, bismuth carbonate, bismuth nitrate, bismuth oxychloride, bismuth subnitrate, bismuth sulfate, bismuth trichloride, cadmium carbonate, cadmium chloride, cadmium iodide, cadmium nitrate, cadmium oxide, Cadmium sulfate, calcium bromide, calcium carbonate, calcium chloride, calcium fluoride, calcium nitrate, calcium oxide powder, calcium sulfate, chromium(III) chloride, chromium(III) nitrate, chromium(III) oxide, chromium piconylate, chromium trioxide, cobalt(II) carbonate, cobalt(II) chloride, cobalt(II) nitrate, cobalt(II) sulfate, cobalt oxide, cupric bromide, cupric carbonate, cupric chloride, cupric nitrate, cupric oxide powder, copper sulfate, cuprous bromide, cuprous chloride, cuprous iodide, cuprous oxide, dysprosium oxide, europium oxide, dichloromethane Iron, ferric nitrate, ferric oxide, ferric sulfate, ferric chloride, ferrous sulfate, gadolinium(III) oxide, germanium dioxide, holmium oxide, indium(III) chloride, iridium trichloride, lanthanum carbonate, lanthanum chloride, lanthanum nitrate, lanthanum oxalate, lanthanum oxide, lead(II) chloride, lead acetate, lead bromide, lead carbonate, lead dioxide, lead monoxide, lead nitrate, lead oxide, lead sulfate, magnesium bromide, magnesium nitrate, magnesium oxide, manganese(II) sulfate, manganese carbonate, manganese dioxide, mercury iodide, mercury nitrate, mercury oxide, molybdenum trioxide, neodymium oxide, carbonate Nickel, nickel chloride, nickel nitrate, nickel oxide, nickel sulfate, niobium pentoxide, platinum chloride, potassium bromide, potassium carbonate, potassium chloride, potassium chloroplatinate, potassium chromate, potassium iodate, potassium iodide, potassium nitrate, potassium pyroantimonate, rhodium trichloride, rubidium chloride, samarium oxide, selenium dioxide, silver benzoate, silver bromide, silver carbonate, silver chloride, silver iodide, silver nitrate, silver oxide, silver sulfate, sodium arsenate, sodium arsenite, sodium bromide, sodium carbonate, sodium fluoride, sodium iodide, sodium metaborate,The composition for immunoassay according to embodiment 1 or embodiment 2, selected from the list consisting of sodium oxalate, sodium silicate, sodium silicofluoride, sodium stannate, sodium tellurite, stannous chloride stannate, stannic oxide, stannous chloride, stannous sulfate, strontium carbonate, titanium tetrachloride, tungsten(VI) oxide, vanadium pentoxide, yttrium oxide, zinc chloride, zinc cyanide, zinc nitrate, zinc oxide, zinc sulfate, zirconium dioxide and zirconyl nitrate.

[0172] 4. The immunoassay composition of any of the preceding embodiments, wherein the buffer compound is at a concentration of about 1 mM to 500 mM.

[0173] 5. The immunoassay composition of any of the preceding embodiments, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 100 mM.

[0174] 6. The immunoassay composition of any of the preceding embodiments, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 10 mM to about 100 mM.

[0175] 7. The immunoassay composition of any of the preceding embodiments, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM.

[0176] 8. The immunoassay composition of any of the preceding embodiments, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM.

[0177] 9. The immunoassay composition of any of the preceding embodiments, wherein the salt is at a concentration of from about 1 mM to about 1000 mM.

[0178] 10. The immunoassay composition of any of the preceding embodiments, comprising sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM.

[0179] 11. The immunoassay composition of any of the preceding embodiments, comprising sodium chloride (NaCl) at a concentration of about 50 to about 500 mM.

[0180] 12. The immunoassay composition of any of the preceding embodiments, comprising sodium chloride (NaCl) at a concentration of about 200 to about 400 mM.

[0181] 13. The immunoassay composition of any of the preceding embodiments, comprising sodium chloride (NaCl) at a concentration of about 100 to about 200 mM.

[0182] 14. The immunoassay composition of any one of embodiments 1 to 11 and 13, comprising sodium chloride (NaCl) at a concentration of about 150 mM.

[0183] 15. The immunoassay composition according to any one of embodiments 1 to 12, comprising sodium chloride (NaCl) at a concentration of about 400 mM.

[0184] 16. The immunoassay composition of any one of embodiments 1 to 12, comprising sodium chloride (NaCl) at a concentration of about 275 mM.

[0185] 17. The immunoassay composition of any of the preceding embodiments, comprising Tris(tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), sodium chloride, and one or more block copolymers.

[0186] 18. The immunoassay composition of any of the preceding embodiments, comprising Tris-Buffered Saline (TBS).

[0187] 19. The immunoassay composition according to embodiment 18, wherein the Tris-buffered saline (TBS) contains Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 100 mM to about 200 mM.

[0188] 20. The immunoassay composition of embodiment 19, wherein the Tris-Buffered Saline (TBS) comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a concentration of about 150 mM.

[0189] 21. The immunoassay composition according to embodiment 18, wherein the Tris-buffered saline (TBS) contains Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 50 mM to about 500 mM.

[0190] 22. The composition for use in an immunoassay according to embodiment 21, wherein the Tris-Buffered Saline (TBS) comprises Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a concentration of about 400 mM.

[0191] 23. The immunoassay composition of any one of embodiments 1 to 13 and 15 to 22, comprising Tris-Buffered Saline (TBS) and additional NaCl, so as to provide a composition comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a total concentration of about 200 mM to about 500 mM.

[0192] 24. The immunoassay composition of embodiment 23, comprising Tris-Buffered Saline (TBS) and additional NaCl to provide a composition comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a total concentration of about 200 mM to about 400 mM.

[0193] 25. The immunoassay composition of embodiment 24, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a total concentration of about 275 mM.

[0194] 26. The immunoassay composition of any of the preceding embodiments, comprising one or more block copolymers in a total concentration of about 0.01% (w / v) to about 2% (w / v).

[0195] 27. The immunoassay composition of any of the preceding embodiments, comprising one or more block copolymers in a total concentration of about 0.25% (w / v) to about 1.5% (w / v).

[0196] 28. The immunoassay composition of any of the preceding embodiments, comprising one or more block copolymers at a total concentration of about 0.05% (w / v).

[0197] 29. The immunoassay composition of any of the preceding embodiments, comprising one or more block copolymers at a total concentration of about 0.125% (w / v).

[0198] 30. The immunoassay composition of any of the preceding embodiments, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM, and one or more block copolymers at a total concentration of about 0.01% (w / v) to about 2% (w / v).

[0199] 3. The immunoassay composition of any of the preceding embodiments, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 31.25 mM, sodium chloride (NaCl) at a concentration of 150 mM, and the block copolymer at a concentration of 0.05% (w / v).

[0200] The immunoassay composition according to any one of embodiments 1 to 30, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 32.25 mM, sodium chloride (NaCl) at a concentration of 275 mM, and a block copolymer at a concentration of 0.125% (w / v).

[0201] 33. A composition for an immunoassay comprising a buffer compound, one or more salts, one or more blocking agents, and one or more block copolymers.

[0202] 34. The composition for use in an immunoassay according to embodiment 33, wherein the one or more blocking agents are selected from the list consisting of proteins, such as phosphoproteins, polymers and small molecules.

[0203] 35. The immunoassay composition of embodiment 34, wherein the protein is a phosphoprotein, optionally wherein the phosphoprotein is casein.

[0204] 36. The immunoassay composition according to embodiment 34, wherein the protein is bovine serum albumin (BSA).

[0205] 37. The composition for use in an immunoassay according to embodiment 34, wherein the polymer is PVP (polyvinylpyrrolidone) or PVA (polyvinyl alcohol).

[0206] 38. The composition for use in an immunoassay according to embodiment 34, wherein the small molecule is ethanolamine.

[0207] 39. The composition for use in an immunoassay according to any one of embodiments 33 to 38, comprising at least two salts, such as 2, 3, 4, 5 or 6 salts.

[0208] 40. The composition for use in an immunoassay according to any one of embodiments 33 to 39, comprising two salts.

[0209] 41. The buffer compound is Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), Bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), Tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid), HEPES (4-(2-hydroxyethyl)amino)acetic acid), or glycine (glycine). 41. The immunoassay composition according to any one of embodiments 33 to 40, wherein the aryl group is selected from the list consisting of (1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)amino]ethanesulfonic acid), (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), (2-(hydroxyethyl)-1-piperazine ...

[0210] 42. One or more salts of the following substances are not to be construed as limiting the scope of the invention: sodium chloride, calcium chloride, aluminum ammonium sulfate, aluminum chloride, aluminum chloride, aluminum fluoride, aluminum nitrate, aluminum potassium sulfate, aluminum sulfate, antimony(III) sulfide, antimony trichloride, barium bromide, barium carbonate, barium chloride, barium chromate, barium nitrate, barium perchlorate, barium sulfate, bismuth carbonate, bismuth nitrate, bismuth oxychloride, bismuth subnitrate, bismuth sulfate, bismuth trichloride, cadmium carbonate, cadmium chloride, cadmium iodide, Cadmium nitrate, cadmium oxide, cadmium sulfate, calcium bromide, calcium carbonate, calcium fluoride, calcium nitrate, calcium oxide powder, calcium sulfate, chromium (III) chloride, chromium (III) nitrate, chromium (III) oxide, chromium piconylate, chromium trioxide, cobalt (II) carbonate, cobalt (II) chloride, cobalt (II) nitrate, cobalt (II) sulfate, cobalt oxide, cupric bromide, cupric carbonate, cupric chloride, cupric nitrate, cupric oxide powder, copper sulfate, cuprous bromide, cuprous chloride, cuprous iodide, cuprous oxide, dysprosium oxide, eutectic oxide Rhodium, ferric chloride, ferric nitrate, ferric oxide, ferric sulfate, ferric chloride, ferrous sulfate, gadolinium(III) oxide, germanium dioxide, holmium oxide, indium(III) chloride, iridium trichloride, lanthanum carbonate, lanthanum chloride, lanthanum nitrate, lanthanum oxalate, lanthanum oxide, lead(II) chloride, lead acetate, lead bromide, lead carbonate, lead dioxide, lead monoxide, lead nitrate, lead oxide, lead sulfate, magnesium bromide, magnesium nitrate, magnesium oxide, manganese(II) sulfate, manganese carbonate, manganese dioxide, mercury iodide, mercury nitrate, mercury oxide, molybdenum trioxide , neodymium oxide, nickel carbonate, nickel chloride, nickel nitrate, nickel oxide, nickel sulfate, niobium pentoxide, platinum chloride, potassium bromide, potassium carbonate, potassium chloride, potassium chloroplatinate, potassium chromate, potassium iodate, potassium iodide, potassium nitrate, potassium pyroantimonate, rhodium trichloride, rubidium chloride, samarium oxide, selenium dioxide, silver benzoate, silver bromide, silver carbonate, silver chloride, silver iodide, silver nitrate, silver oxide, silver sulfate, sodium arsenate, sodium arsenite, sodium bromide, sodium carbonate, sodium fluoride, sodium iodide,The composition for immunoassay according to any one of embodiments 33 to 41, selected from the list consisting of sodium metaborate, sodium oxalate, sodium silicate, sodium silicofluoride, sodium stannate, sodium tellurite, stannous chloride stannate, stannic oxide, stannous chloride, stannous sulfate, strontium carbonate, titanium tetrachloride, tungsten(VI) oxide, vanadium pentoxide, yttrium oxide, zinc chloride, zinc cyanide, zinc nitrate, zinc oxide, zinc sulfate, zirconium dioxide and zirconyl nitrate.

[0211] 43. The composition for immunoassays according to any one of embodiments 33 to 42, wherein the buffer compound is at a concentration of about 1 mM to 500 mM.

[0212] 44. The composition for immunoassay according to any one of embodiments 33 to 43, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 100 mM.

[0213] 45. The composition for immunoassay according to any one of embodiments 33 to 44, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM.

[0214] 46. ​​The immunoassay composition according to any one of embodiments 33 to 45, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM.

[0215] 47. The composition for use in an immunoassay according to any one of embodiments 33 to 46, wherein one of the one or more salts is at a concentration of from about 1 mM to about 1000 mM.

[0216] 48. The composition for immunoassay according to any one of embodiments 33 to 47, comprising sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM.

[0217] 49. The composition for immunoassay according to any one of embodiments 33 to 48, comprising sodium chloride (NaCl) at a concentration of about 200 mM to about 400 mM.

[0218] 50. The composition for immunoassay according to any one of embodiments 33 to 48, comprising sodium chloride (NaCl) at a concentration of about 300 mM to about 500 mM.

[0219] 51. The composition for immunoassay according to any one of embodiments 33 to 48, comprising sodium chloride (NaCl) at a concentration of about 100 mM to about 200 mM.

[0220] 52. The composition for use in an immunoassay according to embodiment 48, comprising sodium chloride (NaCl) at a concentration of about 400 mM.

[0221] 53. The composition for use in an immunoassay according to embodiment 48, comprising sodium chloride (NaCl) at a concentration of about 275 mM.

[0222] 54. The composition for use in an immunoassay according to embodiment 48, comprising sodium chloride (NaCl) at a concentration of about 150 mM.

[0223] 55. The composition for use in an immunoassay according to any one of embodiments 33 to 54, wherein one of the one or more salts is at a concentration of from about 1 mM to about 200 mM.

[0224] 56. A composition for immunoassays according to any one of embodiments 33 to 55, comprising calcium chloride (CaCl2) at a concentration of about 1 mM to about 200 mM.

[0225] 57. A composition for immunoassays according to any one of embodiments 33 to 56, comprising calcium chloride (CaCl2) at a concentration of about 5 mM to about 20 mM.

[0226] 58. The composition for immunoassay according to any one of embodiments 33 to 57, comprising calcium chloride (CaCl2) at a concentration of about 10 mM to about 20 mM.

[0227] 59. The composition for immunoassay described in embodiment 58, comprising calcium chloride (CaCl2) at a concentration of about 16 mM.

[0228] 60. The composition for use in an immunoassay according to embodiment 57, comprising calcium chloride (CaCl2) at a concentration of about 8 mM.

[0229] 61. The composition for immunoassay according to any one of embodiments 33 to 60, comprising Tris-Buffered Saline (TBS).

[0230] 62. The composition for immunoassay according to any one of embodiments 33 to 61, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 100 mM to 200 mM.

[0231] 63. The composition for use in an immunoassay according to embodiment 62, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a concentration of about 150 mM.

[0232] 64. The composition for immunoassay according to any one of embodiments 33 to 61, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM and NaCl at a concentration of about 200 mM to 400 mM.

[0233] 65. The composition for use in an immunoassay according to embodiment 64, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM and NaCl at a concentration of about 275 mM.

[0234] 66. The composition for immunoassay according to any one of embodiments 33 to 61, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM, NaCl at a concentration of about 200 mM to about 400 mM, and CaCl2 at a concentration of about 5 mM to about 20 mM.

[0235] 67. The composition for use in an immunoassay according to embodiment 66, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, NaCl at a concentration of about 275 mM, and CaCl2 at a concentration of about 8 mM.

[0236] 68. The composition for immunoassay according to any one of embodiments 33 to 61, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 20 mM to about 30 mM, NaCl at a concentration of about 300 mM to about 500 mM, and CaCl2 at a concentration of about 10 mM to about 20 mM.

[0237] 69. The composition for use in an immunoassay according to embodiment 68, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, NaCl at a concentration of about 400 mM, and CaCl2 at a concentration of about 16 mM.

[0238] 70. The composition for use in an immunoassay according to any one of embodiments 33 to 69, comprising one or more block copolymers in a total concentration of about 0.05% (w / v) to about 5% (w / v).

[0239] 71. The composition for use in an immunoassay according to any one of embodiments 33 to 70, comprising one or more block copolymers at a total concentration of from about 0.75% (w / v) to about 1.5% (w / v).

[0240] 72. The immunoassay composition according to any one of embodiments 33 to 71, comprising one or more block copolymers at a total concentration of about 1% (w / v).

[0241] 73. A composition for an immunoassay according to any one of embodiments 33 to 72, comprising one block copolymer at a concentration of about 1% (w / v).

[0242] 74. The composition for use in an immunoassay according to any one of embodiments 33 to 70, comprising one or more block copolymers in a total concentration of about 0.05% (w / v) to about 0.5% (w / v).

[0243] 75. The composition for use in an immunoassay according to embodiment 74, comprising one or more block copolymers at a total concentration of about 0.25% (w / v).

[0244] 76. The composition for use in an immunoassay according to embodiment 75, comprising one block copolymer at a concentration of about 0.25% (w / v).

[0245] 77. The composition for use in an immunoassay according to embodiment 70, comprising one or more block copolymers at a total concentration of about 0.125% (w / v).

[0246] 78. The composition for use in an immunoassay according to embodiment 77, comprising one block copolymer at a concentration of about 0.125% (w / v).

[0247] 79. A total concentration of about 0.05% (w / v) to about 10% (w / v), for example, about 0.05% (w / v) to about 9% (w / v), about 0.05% (w / v) to about 8% (w / v), about 0.05% (w / v) to about 7% (w / v), about 0.05% (w / v) to about 6% (w / v), about 0.05% (w / v) to about 5% (w / v), about 0.05% (w / v) to about 4% (w / v), about 0.05% (w / v) to about 3% (w / v), about 0.05% (w / v) to about 2.5% (w / v), about 0.05% (w / v) to about 2% 79. The immunoassay composition according to any one of embodiments 33 to 78, comprising one or more blocking agents at a total concentration of about 0.05% (w / v) to about 1.5% (w / v), about 0.05% (w / v) to about 1% (w / v), about 0.05% (w / v) to about 0.5% (w / v), about 0.05% (w / v) to about 0.4% (w / v), about 0.05% (w / v) to about 0.3% (w / v), about 0.05% (w / v) to about 0.2% (w / v), or about 0.05% (w / v) to about 0.1% (w / v).

[0248] 80. The composition for use in an immunoassay according to embodiment 79, comprising a blocking agent comprising bovine serum albumin at a concentration of about 0.05% (w / v) to about 10% (w / v).

[0249] 81. The composition for use in an immunoassay according to embodiment 80, comprising a blocking agent comprising bovine serum albumin at a concentration of about 0.5% (w / v).

[0250] 82. The composition for use in an immunoassay according to embodiment 80, comprising a blocking agent comprising bovine serum albumin at a concentration of about 2.5% (w / v).

[0251] 83. The immunoassay composition according to any one of embodiments 33 to 82, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM, calcium chloride (CaCl2) at a concentration of about 1 mM to 200 mM, bovine serum albumin (BSA) at a concentration of about 0.05% (w / v) to about 10% (w / v), and one or more block copolymers at a total concentration of about 0.05% (w / v) to about 5% (w / v).

[0252] The immunoassay composition of embodiment 80, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 84.25 mM, sodium chloride (NaCl) at a concentration of 400 mM, calcium chloride (CaCl2) at a concentration of 16 mM, bovine serum albumin (BSA) at a concentration of 5% (w / v), and a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 0.25% (w / v).

[0253] The immunoassay composition of embodiment 80, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of 85.25 mM, sodium chloride (NaCl) at a concentration of 275 mM, calcium chloride (CaCl2) at a concentration of 8 mM, bovine serum albumin (BSA) at a concentration of 2.5% (w / v), and a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 0.125% (w / v).

[0254] 86. A composition for immunoassays according to any one of embodiments 33 to 79, comprising casein at a concentration of about 0.1% (w / v) to about 3% (w / v).

[0255] 87. The composition for use in an immunoassay according to embodiment 86, comprising casein at a concentration of about 0.4% (w / v) to about 0.6% (w / v).

[0256] 88. The composition for use in an immunoassay according to embodiment 86 or embodiment 87, comprising casein at a concentration of about 0.5% (w / v).

[0257] 89. The composition for immunoassay according to any one of embodiments 33 to 88, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), sodium chloride (NaCl), calcium chloride (CaCl2), casein, and one or more block copolymers.

[0258] 90. The composition for use in an immunoassay according to embodiment 89, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to 1000 mM, calcium chloride (CaCl2) at a concentration of about 1 mM to 200 mM, casein at a concentration of about 0.1% (w / v) to about 2% (w / v), and one or more block copolymers at a total concentration of about 0.05% (w / v) to about 5% (w / v).

[0259] 91. The immunoassay composition of embodiment 90, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, sodium chloride (NaCl) at a concentration of about 150 mM, calcium chloride (CaCl2) at a concentration of about 16 mM, casein at a concentration of about 0.5% (w / v), and a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 1% (w / v).

[0260] 92. The immunoassay composition of any of the preceding embodiments, wherein the block copolymer is a polyoxyethylene-polyoxypropylene block copolymer.

[0261] 93. A polyoxyethylene-polyoxypropylene block copolymer having the formula I

[0262] [ka]

[0263] Formula I (wherein x and z are 2 to 130, and b is 15 to 67).

[0264] 94. The composition for immunoassay according to embodiment 93, wherein x and z=120-130, and b=50-60.

[0265] 95. The immunoassay composition of any of the previous embodiments, wherein the block copolymer is Synperonic® F108 having the following formula:

[0266] [ka]

[0267] 96. The immunoassay composition of any of the preceding embodiments, further comprising one or more chaotropic agents.

[0268] 97. The immunoassay composition of any of the preceding embodiments, further comprising one or more denaturing agents.

[0269] 98. The immunoassay composition of any of the preceding embodiments, further comprising one or more chaotropic agents and denaturing agents.

[0270] 99. The composition for immunoassay according to any one of embodiments 96 to 98, wherein the one or more chaotropic agents and / or denaturing agents are selected from the list consisting of guanidine salts, urea or betaine.

[0271] 100. The immunoassay composition of embodiment 99, wherein the one or more chaotropic and / or denaturing agents is guanidine hydrochloride.

[0272] 101. The composition for immunoassay according to any one of embodiments 96 to 100, wherein the one or more chaotropic agents and / or denaturing agents are at a concentration of about 1 mM to about 1000 mM.

[0273] 102. The immunoassay composition according to any one of embodiments 96 to 101, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM, calcium chloride (CaCl2) at a concentration of about 1 mM to about 200 mM, casein at a concentration of about 0.1% (w / v) to about 2% (w / v), one or more block copolymers at a total concentration of about 0.05% (w / v) to about 5% (w / v), and one or more chaotropic agents and / or denaturants at a concentration of about 1 mM to about 1000 mM.

[0274] 103. The immunoassay composition of embodiment 102, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, sodium chloride (NaCl) at a concentration of about 150 mM, calcium chloride (CaCl2) at a concentration of about 16 mM, casein at a concentration of about 0.5% (w / v), a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 1% (w / v), and guanidine hydrochloride (guanidine HCl) at a concentration of about 200 mM.

[0275] 104. The immunoassay composition according to any one of embodiments 96 to 101, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 1 mM to about 500 mM, sodium chloride (NaCl) at a concentration of about 1 mM to about 1000 mM, calcium chloride (CaCl2) at a concentration of about 1 mM to about 200 mM, bovine serum albumin at a concentration of about 0.5% (w / v) to about 10% (w / v), one or more block copolymers at a total concentration of about 0.05% (w / v) to about 5% (w / v), and one or more chaotropic agents and / or denaturants at a concentration of about 1 mM to about 1000 mM.

[0276] 105. The immunoassay composition of embodiment 104, comprising Tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol) at a concentration of about 25 mM, sodium chloride (NaCl) at a concentration of about 275 mM, calcium chloride (CaCl2) at a concentration of about 8 mM, bovine serum albumin at a concentration of about 2.5% (w / v), a polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) at a concentration of about 0.125% (w / v), and guanidine hydrochloride (guanidine HCl) at a concentration of about 300 mM.

[0277] 106. A composition for an immunoassay comprising one or more chaotropic and / or denaturing agents.

[0278] 107. The composition for use in an immunoassay according to embodiment 106, wherein the one or more chaotropic and / or denaturing agents are selected from the list consisting of guanidine salts, urea or betaine.

[0279] 108. The immunoassay composition according to embodiment 107, wherein the one or more chaotropic and / or denaturing agents is guanidine hydrochloride.

[0280] 109. The composition for immunoassays according to any one of embodiments 106 to 108, wherein the one or more chaotropic agents and / or denaturing agents are at a concentration of about 1 mM to about 1000 mM.

[0281] 110. The composition for an immunoassay according to any one of embodiments 106 to 109, further comprising a proteinaceous matrix, optionally comprising bovine serum albumin (BSA) and / or casein.

[0282] 111. The immunoassay composition of any one of the preceding embodiments, further comprising a biocide.

[0283] 112. The immunoassay composition of embodiment 111, wherein the biocide comprises 2-methyl-4-isothiazolin-3-one (MIT), 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT) or a mixture thereof (e.g., a ProClin™ biocide such as ProClin™ 300).

[0284] 113. The composition for use in an immunoassay according to embodiment 111 or 112, comprising a biocide at a concentration of about 0.01% (w / v) to about 2% (w / v), for example, about 0.01% (w / v) to about 1% (w / v), about 0.01% (w / v) to about 0.5% (w / v), about 0.01% (w / v) to about 0.1% (w / v), or about 0.02% (w / v) to about 0.08% (w / v).

[0285] 114. The composition for use in an immunoassay according to embodiment 113, comprising a biocide at a concentration of about 0.03% (w / v) to about 0.07% (w / v) or about 0.05% (w / v).

[0286] 115. The composition for immunoassay of any one of the preceding embodiments, further comprising an antifoaming agent. 116. The composition for use in an immunoassay according to embodiment 115, wherein the antifoaming agent comprises a non-silicone antifoaming agent in a polyol-based dispersion (e.g., Antifoam 204).

[0287] 117. The composition for immunoassay according to embodiment 115 or 116, comprising an antifoaming agent at a concentration of about 0.01% (w / v) to about 2% (w / v), for example, about 0.01% (w / v) to about 1% (w / v), about 0.01% (w / v) to about 0.5% (w / v), about 0.01% (w / v) to about 0.1% (w / v), or about 0.02% (w / v) to about 0.08% (w / v).

[0288] 118. The composition for use in an immunoassay according to embodiment 117, comprising an antifoaming agent at a concentration of about 0.03% (w / v) to about 0.07% (w / v) or about 0.05% (w / v).

[0289] 119. The immunoassay composition of any one of the preceding embodiments, further comprising a heterophilic blocking composition.

[0290] 120. The immunoassay composition of any one of the preceding embodiments, which is a liquid, a gel or an emulsion.

[0291] 121. The immunoassay composition of any one of the preceding embodiments, comprising water.

[0292] 122. The composition for immunoassay of any one of the preceding embodiments, which is a liquid composition comprising the components of the composition in an aqueous solution, a suspension, or a combination thereof.

[0293] 123. A composition for immunoassays according to any one of embodiments 1 to 119, which is lyophilized.

[0294] 124. A method for preparing a composition for an immunoassay according to any one of embodiments 1 to 122, comprising mixing water with an additional component of the composition to obtain the composition.

[0295] 125. The method according to embodiment 124, comprising mixing an aqueous buffered solution or water with an additional component of the composition to obtain the composition.

[0296] 126. The method of embodiment 124 or 125, wherein the additional component is provided in solid form, as an aqueous solution, or as a combination thereof.

[0297] 127. The method of embodiment 126, wherein one or more of the additional ingredients are provided in lyophilized form prior to mixing with the aqueous buffer solution or water.

[0298] 128. A method for preparing a composition for an immunoassay according to any one of embodiments 124 to 127, wherein the composition comprises a blocking agent, the method comprising obtaining a liquid composition comprising a buffer compound, a salt, one or more block copolymers and optionally one or more additional components of the composition, providing a lyophilized blocking agent, and mixing the liquid composition with the lyophilized blocking agent to obtain the composition for an immunoassay.

[0299] 129. The method of embodiment 128, wherein the composition further comprises one or more chaotropic agents and / or denaturing agents, and further comprising providing the one or more chaotropic agents and / or denaturing agents in lyophilized form, and mixing the liquid composition with the lyophilized blocking agent and the lyophilized one or more chaotropic agents and / or denaturing agents to obtain a composition for immunoassay.

[0300] 130. The method of embodiment 128 or 129, wherein the blocking agent is BSA. 131. The method of any one of embodiments 129 or 130, wherein the one or more chaotropic agents and / or denaturing agents are guanidine salts, such as guanidine hydrochloride.

[0301] 132. The method according to any one of embodiments 124 to 131, wherein the components of the composition are provided to prepare a composition having the components and / or concentrations of any one of embodiments 1 to 122.

[0302] 133. A method for performing an immunoassay, comprising contacting a biological sample with one or more compositions according to any one of embodiments 1 to 122.

[0303] 134. A method of performing an immunoassay according to embodiment 133, which is for detecting the presence of an analyte of interest in a biological sample.

[0304] 135. A method for performing an immunoassay according to embodiments 133-134, in which the biological sample is contacted simultaneously or sequentially with one or more compositions according to any one of embodiments 1-122.

[0305] 136. A method for performing an immunoassay according to any one of embodiments 133 to 135, in which one or more compositions are used as a diluent for a biological sample.

[0306] 137. A method for performing an immunoassay according to any one of embodiments 133 to 136, comprising contacting the biological sample with a capture antibody, wherein one or more compositions are used as a capture diluent when contacting the biological sample with the capture antibody.

[0307] 138. A method for performing an immunoassay according to any one of embodiments 133 to 137, comprising adding a detection antibody, wherein when adding the detection antibody, one or more compositions are used as a detection diluent.

[0308] 139. A method of performing an immunoassay according to embodiment 138, wherein adding the detection antibody can be performed simultaneously with or after contacting the biological sample with the capture antibody.

[0309] 140. A method for performing an immunoassay according to embodiment 138 or 139, in which contacting the biological sample with the capture antibody is followed by a washing step using a washing solution, and adding a detection diluent to the capture antibody prior to or simultaneously with the addition of the detection antibody.

[0310] 141. A method for performing an immunoassay according to any one of embodiments 133 to 140, wherein one or more compositions are used as a washing solution.

[0311] 142. A method for performing an immunoassay according to embodiment 141, wherein the wash solution comprises or consists essentially of Tris-buffered saline and a block copolymer, as defined in any of the preceding embodiments.

[0312] 143. A method for carrying out an immunoassay according to any one of embodiments 133 to 142, wherein one or more of the compositions is used as a buffer.

[0313] 144. The method of any one of embodiments 133 to 143, wherein the biological sample is a blood sample, such as a whole blood sample or a blood fraction, amniotic fluid, aqueous humor, bile, plasma, breast milk, cerebrospinal fluid (CSF), endolymph, extracellular fluid, exudate, gastric acid, hemolymph, interstitial fluid, lymph, mucus, pericardial fluid, peritoneal fluid, sweat (perspiration), sputum, pus, saliva, semen, synovial fluid, tears, urine, vaginal fluid, vomit, saliva, other biological fluids or a swab sample.

[0314] 145. The method according to any one of embodiments 133 to 144, wherein the biological sample is a whole blood sample.

[0315] 146. The method of any one of embodiments 133-145, wherein the biological sample comprises an analyte of interest.

[0316] 147. The method of any one of embodiments 134 to 146, wherein the analyte of interest is a protein.

[0317] 148. The method of embodiment 147, wherein the analyte of interest is brain natriuretic peptide or N-terminal proBNP.

[0318] 149. The method of embodiment 147, wherein the analyte of interest is cardiac troponin or cardiac troponin subunit I (cTnI).

[0319] 150. The method according to any one of embodiments 133 to 149, wherein the immunoassay is a competitive sandwich immunoassay.

[0320] 151. The method of any one of embodiments 133-150, wherein the immunoassay comprises obtaining electrochemical measurements to detect the analyte of interest.

[0321] 152. The method of any one of embodiments 133-151, wherein the electrochemical measurements are amperometric, voltametric, potentiometric, impedancemetric, or electrochemical impedance spectroscopy measurements.

[0322] 153. The method of embodiment 152, wherein the electrochemical measurements are chronoamperometric measurements.

[0323] 154. The method is for measuring an analyte of interest in a biological sample, a. A biological sample is a first antibody or antigen-binding portion thereof capable of binding to the analyte of interest; a second antibody or antigen-binding portion thereof capable of binding to the analyte of interest conjugated to an enzyme; and An assay diluent, the composition being any one of embodiments 1 to 122. to form an assay mixture; b. contacting the assay mixture with a substrate for the enzyme, where the enzyme substrate is converted by the enzyme into an electroactive molecule; and c. Obtaining an electrochemical measurement to measure the analyte of interest.

[0324] 155. In some embodiments of the method, the method is for measuring an analyte of interest in a biological sample, a. The biological sample is a magnetically susceptible bead conjugated to a first antibody or an antigen-binding portion thereof capable of binding to the analyte of interest; a second antibody or antigen-binding portion thereof capable of binding to the analyte of interest conjugated to an enzyme; and An assay diluent, the composition being any one of embodiments 1 to 122. to form an assay mixture; b. holding magnetically susceptible beads on electrodes using a magnetic field; c. contacting the magnetically susceptible beads with a substrate of the enzyme, where the enzyme substrate is converted by the enzyme into an electroactive molecule; and d. Obtaining an electrochemical measurement using the electrode. The method of any one of embodiments 133-154, comprising:

[0325] 156. In some embodiments of the method, the method is for measuring an analyte of interest in a biological sample, a. mixing said biological sample with magnetically susceptible beads conjugated to a first antibody or antigen-binding portion thereof capable of binding to said analyte of interest in the presence of a capture diluent, wherein the capture diluent is a composition according to any one of embodiments 1-122; b. immobilizing magnetically susceptible beads on a magnet and washing the beads; c. providing a detection diluent with a second antibody or antigen-binding portion thereof capable of binding to the analyte of interest conjugated to an enzyme to produce a detection solution, wherein the detection diluent is a composition according to any one of embodiments 1 to 122; d. adding detection solution to the washed magnetically susceptible beads; e. Washing the magnetically susceptible beads; f. contacting the magnetically susceptible bead with a substrate of the enzyme, whereby the enzyme substrate is converted by the enzyme into an electroactive molecule; and g. Obtaining an electrochemical measurement to measure the analyte of interest.

[0326] 157. The method of embodiment 156, wherein the washing steps b and e are carried out using a washing solution, optionally wherein the washing solution is a composition described in any one of embodiments 1 to 122.

[0327] 158. The method of embodiment 157, wherein the washing solution comprises or consists essentially of Tris-buffered saline and a block copolymer, as defined in any of the preceding embodiments.

[0328] 159. The method according to any one of embodiments 137 to 158, wherein the capture diluent is a composition according to any one of embodiments 1 to 122, comprising a buffer compound, at least one salt, one or more block copolymers, one or more blocking agents, and one or more chaotropic agents and / or denaturing agents.

[0329] 160. The method of embodiment 159, wherein the capture diluent comprises at least two salts. 161. The method of embodiment 159 or 160, wherein the capture diluent comprises Tris, NaCl, CaCl2, polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108), bovine serum albumin and guanidine hydrochloride (guanidine HCl).

[0330] 162. The method according to any one of embodiments 138 to 161, wherein the detection diluent is a composition according to any one of embodiments 1 to 122, comprising a buffer compound, at least one salt, one or more block copolymers, and one or more blocking agents.

[0331] 163. The method of embodiment 162, wherein the detection diluent comprises at least two salts. 164. The method of embodiment 162 or 163, wherein the detection diluent comprises Tris, NaCl, CaCl2, polyoxyethylene-polyoxypropylene block copolymer (e.g., Synperonic® F108) and bovine serum albumin.

[0332] 165. The method according to any one of embodiments 133 to 164, wherein one or more compositions according to any one of embodiments 1 to 122 are prepared by a method according to any one of embodiments 124 to 132.

[0333] 166. The method of any one of embodiments 133 to 165, wherein the immunoassay is performed in a microfluidic cartridge, optionally a microfluidic disposable cartridge.

[0334] 167. The method according to embodiment 166, wherein one or more compositions according to any one of embodiments 1 to 122 are prepared using a microfluidic cartridge by a method according to any one of embodiments 124 to 132.

[0335] 168. The method according to embodiment 167, wherein the one or more compositions prepared by the method according to any one of embodiments 124 to 132 using a microfluidic cartridge are a capture diluent and / or a detection diluent.

[0336] 169. Use of a composition according to any one of embodiments 1 to 123 in a point-of-care diagnostic assay, such as an immunoassay.

[0337] 170. Use of a composition according to any one of embodiments 1 to 123 in an immunoassay.

[0338] 171. The use according to embodiment 169 or 170, wherein the immunoassay is a competitive sandwich immunoassay.

[0339] 172. The use according to any one of embodiments 169 to 171, wherein the immunoassay comprises obtaining electrochemical measurements to detect the analyte of interest.

[0340] 173. The use according to embodiment 172, wherein the electrochemical measurement is an amperometric, voltametric, potentiometric, impedancemetric, or electrochemical impedance spectroscopy measurement.

[0341] 174. The use according to embodiment 173, wherein the electrochemical measurements are chronoamperometric measurements.

[0342] 175. One or more compositions according to any one of embodiments 1 to 123; Point-of-care immunoassay diagnostic tests and kits comprising:

[0343] 176. In some embodiments, the kit further comprises an instructional material, optionally comprising instructions for carrying out a method of carrying out an immunoassay described in any one of embodiments 133 to 168, the kit described in embodiment 175.

[0344] References [1] HH Bearat, BL Vernon, 11-Environmentally responsive injectable materials, Injectable Biomaterials, 2011, pp. 263-297, ISBN 9781845695880 (incorporated herein by reference)

Claims

1. A composition for immunoassay, comprising a buffering compound, one or more salts, one or more blocking agents, one or more block copolymers, one or more block copolymers, and one or more chaotropic agents and / or denaturing agents.

2. The composition for immunoassay according to claim 1, wherein the one or more chaotropic agents and / or the denaturing agent is guanidine hydrochloride.

3. The composition for immunoassay according to claim 1, wherein the one or more chaotropic agents are at a concentration of 1 mM to 1000 mM.

4. The composition for immunoassay according to claim 1, wherein the one or more chaotropic agents are at a concentration of 1 mM to 500 mM.

5. The one or more blocking agents are proteins, a. the protein is a phosphoprotein, and optionally, the phosphoprotein is casein, or b. the protein is bovine serum albumin (BSA), The composition for immunoassay according to claim 1.

6. The buffering compound is at a concentration of 1 mM to 500 mM, and optionally, the buffering compound is Tris containing Tris (hydroxymethyl) aminomethane or 2-amino-2-(hydroxymethyl) propane-1,3-diol at a concentration of 1 mM to 100 mM. The composition for immunoassay according to claim 1.

7. The composition for immunoassay according to claim 6, comprising Tris (hydroxymethyl) aminomethane or 2-amino-2-(hydroxymethyl) propane-1,3-diol at a concentration of about 1 mM to about 100 mM.

8. One of the one or more salts is at a concentration of about 1 mM to about 1000 mM, and optionally, a. at a concentration of about 300 mM to about 500 mM, or b. at a concentration of about 100 mM to about 200 mM and contains sodium chloride (NaCl). The composition for immunoassay according to claim 1.

9. The composition for immunoassay according to claim 1, further comprising calcium chloride (CaCl ２ ) at a concentration of about 1 mM to about 200 mM.

10. The composition for immunoassay according to claim 5, wherein the bovine serum albumin is at a concentration of about 0.05% (w / v) to about 10% (w / v).

11. a) A blocking agent containing tris (tris (hydroxymethyl) aminomethane or 2-amino-2- (hydroxymethyl) propane-1,3-diol) at a concentration of about 1 mM to about 100 mM, sodium chloride (NaCl) at a concentration of about 100 mM to about 800 mM, and bovine serum albumin at a concentration of about 0.05% (w / v) to about 10% (w / v), and one or more block copolymers at a total concentration of about 0.01% (w / v) to about 1% (w / v), or b) The composition for immunoassay according to claim 1, wherein the concentration of tris (tris (hydroxymethyl) aminomethane or 2-amino-2- (hydroxymethyl) propane-1,3-diol) is about 1 mM to about 100 mM, the concentration of sodium chloride (NaCl) is about 1 mM to about 500 mM, the concentration of casein is about 0.1% (w / v) to about 3% (w / v), and the total concentration of one or more block copolymers is about 0.05% (w / v) to about 2% (w / v).

12. The composition for immunoassay according to claim 5, wherein the casein is about 0.1% (w / v) to about 3% (w / v).

13. The composition for immunoassay according to claim 1, wherein the block copolymer is a polyoxyethylene-polyoxypropylene block copolymer.

14. An antifoaming agent, optionally, the antifoaming agent contains a non-silicone antifoaming agent in a polyol-based dispersion, said antifoaming agent, and / or The composition for immunoassay according to claim 1, further comprising a heterologous affinity blocking composition.

15. Use of a composition containing a buffer compound, one or more salts, one or more blocking agents, one or more block copolymers, one or more block copolymers, one or more chaotropic agents and / or denaturing agents in an immunoassay.

16. The use according to claim 15, wherein the immunoassay comprises obtaining an electrochemical measurement value for detecting an analyte of interest.

17. The use according to claim 16, wherein the electrochemical measurement value is an amperometric, voltammetric, potentiometric, impedance metric, or electrochemical impedance spectroscopy measurement value, optionally the electrochemical measurement value is a chronoamperometry measurement value. A method for preparing a composition comprising a buffering compound, one or more salts, one or more blocking agents, one or more block copolymers, and one or more chaotropic agents and / or denaturing agents, said method comprising: obtaining a liquid composition comprising a buffering compound, a salt, and one or more block copolymers; providing the lyophilized blocking agent and one or more chaotropic agents and / or denaturing agents in lyophilized form; mixing the liquid composition with the lyophilized blocking agent and the lyophilized one or more chaotropic agents and / or denaturing agents to obtain the composition for immunoassay. A method for performing an immunoassay for detecting the presence of an analyte of interest in a biological sample, said method comprising: contacting a composition comprising a buffering compound, one or more salts, one or more blocking agents, one or more block copolymers, and one or more chaotropic agents and / or denaturing agents with the biological sample, wherein the composition is used as a diluent for the biological sample and / or wherein the composition is used as a capture diluent when contacting the biological sample with a capture antibody. The method according to claim 19, comprising obtaining an electrochemical measurement value for detecting the analyte of interest. A method for performing an immunoassay for detecting an analyte of interest in a biological sample, said method comprising performing the immunoassay using a composition comprising a buffering compound, one or more salts, one or more blocking agents, one or more block copolymers, and one or more chaotropic agents and / or denaturing agents.

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