CN113203863A - Buffer solution suitable for interleukin-6 detection - Google Patents
Buffer solution suitable for interleukin-6 detection Download PDFInfo
- Publication number
- CN113203863A CN113203863A CN202110465801.5A CN202110465801A CN113203863A CN 113203863 A CN113203863 A CN 113203863A CN 202110465801 A CN202110465801 A CN 202110465801A CN 113203863 A CN113203863 A CN 113203863A
- Authority
- CN
- China
- Prior art keywords
- buffer
- antibody
- bovine serum
- solution
- kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
- G01N33/6869—Interleukin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/581—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/52—Assays involving cytokines
- G01N2333/54—Interleukins [IL]
- G01N2333/5412—IL-6
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/26—Infectious diseases, e.g. generalised sepsis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the field of detection of interleukin-6 (IL-6), and particularly relates to a buffer solution suitable for detecting interleukin-6, and provides a suitable buffer solution system which comprises 12-18 parts of tris (hydroxymethyl) aminomethane, 30-50 parts of bovine serum albumin, 20-30 parts of glycine, 30-50 parts of trehalose and 20-30 parts of casein. When the system is used for IL-6 detection, the anti-interference capability is strong, the detection result is stable, the repeatability is good, and the detection sensitivity of the system to IL-6 reaches 5 multiplied by 10‑12g/mL or less.
Description
Technical Field
The invention relates to the field of detection of interleukin-6 (IL-6), in particular to a buffer solution suitable for detecting interleukin-6.
Background
IL-6 is a pleiotropic cytokine with wide functions, belonging to one of interleukins. It can be produced by a variety of cells. Interleukin 1(IL-1), tumor necrosis factor-alpha (TNF-alpha), platelet derived factor (PDGF), viral infections, double stranded RNA and c AMP, all induce IL-6 production in normal cells. IL-6 is capable of stimulating the proliferation, differentiation and enhancing the function of cells involved in immune responses. The human IL-6 gene is located on chromosome 7, the molecular weight is 21-30 KD, and the difference is caused by the glycosylation and the phosphate degree of the peptide chain.
IL-6 is a sensitive indicator of early diagnosis of acute infections. When infection and inflammation occur, IL-6 is produced first and increases rapidly, with the level of increase consistent with the severity of the infection, and induces an increase in Procalcitonin (PCT) and C-reactive protein (CRP) following infection. IL-6 can be used as a differential diagnosis index for the early infection of patients and has important reference significance for prognosis. IL-6 can be expressed at a high level for a long time in critically ill patients, can be used as a sensitive index for evaluating the severity and prognosis of patients with sepsis and Multiple Organ Dysfunction Syndrome (MODS), and can reflect the effect of antibiotic treatment more quickly.
In general, in healthy conditions, the plasma IL-6 concentration of newborn infants is 18-26pg/mL, and gradually decreases with age, and the plasma IL-6 concentration of the newborn infants is lower than 10pg/mL when the newborn infants grow into normal adults, although in some inflammatory diseases, the content of IL-6 is increased, but the low total content causes the detection difficulty, and it is necessary to provide a sensitive and accurate detection method of IL-6 content.
Chinese patent application CN109030829A discloses a quantitative kit for detecting canine IL-6 by a homogeneous chemiluminescence method and a using method thereof; the detection solution adopted by the kit comprises a DNA1-IL-6 antibody 1 conjugate, a DNA2-IL-6 antibody 2 conjugate, DNA3 for marking Acridinium Ester (AE), restriction enzyme and graphene oxide, wherein the DNA1 and the DNA2 have 6 base pairs, the DNA3 contains 2 enzyme cutting sites, and has 9 base pairs which are complementary with the DNA1 and the DNA2 respectively, the DNA3 is adsorbed on the surface of the graphene oxide through pi-pi accumulation, and chemiluminescence of the AE marked at the tail end of the graphene oxide is quenched due to CRET; the method is a homogeneous immunoassay method, is simple to operate, greatly shortens the analysis time, can finish the determination of a single sample within 5-10 minutes, and induces the switch of a graphene quenching mechanism through immune reaction by combining an ortho-position touch effect and a chemiluminescent molecular beacon, so that a chemiluminescent signal is released, and washing, separation and purification steps are not required.
Chinese patent application CN106124769A discloses a one-step homogeneous IL-6 detection kit, mainly comprising: anti-IL-6 antibody coupled luminescent microspheres, anti-IL-6 antibody coupled photosensitive microspheres, analysis buffer solution and reaction holes; the invention also discloses a preparation method of the one-step homogeneous phase IL-6 detection kit, which comprises the following steps: preparing an anti-IL-6 antibody coupled luminescent microsphere; preparing an anti-IL-6 antibody coupling photosensitive microsphere and preparing an analysis buffer solution; finally, the invention discloses a using method of the one-step homogeneous phase IL-6 detection kit; the kit has the characteristics of high sensitivity, good specificity, wide detection range, good repeatability, simplicity in operation, no need of cleaning and the like, can be applied to monitoring infection, can improve the accuracy of sepsis diagnosis, and is convenient for clinical detection and use.
Chinese patent application CN107402308A discloses an IL-6 quantitative detection kit and a preparation method thereof. Wherein, the IL-6 quantitative detection kit comprises: a magnetic microsphere reagent coupled with IL-6 antibody, a reaction diluent reagent and an IL-6 calibrator. The detection kit can be used for detecting serum samples on various existing biochemical analyzers, is simple and rapid to operate, only needs about 10-15 minutes to provide detection results, is convenient for early diagnosis of inflammation and sepsis, and is more suitable for popularization and application in various primary medical institutions.
During the use of the IL-6 kit, the detection stability of the standard substance and the quality control substance is very verified on the influence of the result, so that the standard curve of the IL-6 kit is constructed by using a proper buffer solution.
Disclosure of Invention
The invention aims to provide a standard substance buffer solution of IL-6 aiming at the difficulty and low accuracy of IL-6 detection in the prior art.
The purpose of the invention is realized by the following technical scheme.
A buffer solution suitable for detecting interleukin-6 comprises the following components,
12-18 parts of trihydroxymethyl aminomethane, 30-50 parts of bovine serum albumin, 20-30 parts of glycine, 30-50 parts of trehalose and 20-30 parts of casein.
Further, the buffer solution comprises the following components according to the volume of 1000mL,
12-18g of tris (hydroxymethyl) aminomethane, 30-50g of bovine serum albumin, 20-30g of glycine, 30-50g of trehalose, 20-30g of casein, IL-6 recombinant protein and the balance of water, wherein the concentration of the IL-6 recombinant protein is 10-500 pg/mL.
Further, the buffer pH was 7.2-8.8.
Further, the pH of the buffer is adjusted using sodium hydroxide or hydrochloric acid.
The application of the buffer solution is the application in preparing an IL-6 detection product.
A kit comprising the buffer, comprising the following components:
reagent A: alkaline phosphatase-IL-6 antibody;
and (3) reagent B: biotin-IL-6 antibodies;
and (3) reagent C: streptavidin-magnetic microparticles;
and three different concentrations of said buffer, wherein the three different concentrations of said buffer differ only in the amount of IL-6 recombinant protein.
Further, in the three buffers with different concentrations, the concentration of the IL-6 is 110-15 pg/mL of the standard, 310pg/mL of the standard 2290-.
Further, the specific composition of the reagent A is a mixture of alkaline phosphatase-IL-6 antibody and buffer 8;
the buffer solution 8 comprises, based on 1000mL total weight, 3.5-7g of tris (hydroxymethyl) aminomethane, 7-9g of sodium chloride, 30-50g of bovine serum albumin, 80-100mL of bovine serum, 10-20mL of sheep serum, 10-20mL of horse serum, and the balance of water, wherein the pH value is 5.8-7.6, and the content of alkaline phosphatase-IL-6 antibody is 2-4. mu.g/mL, preferably 2. mu.g/mL.
Further, the pH of the buffer is adjusted using sodium hydroxide or hydrochloric acid.
Further, the specific composition of the reagent B is a mixture of biotin-IL-6 antibody and buffer 8;
the buffer solution 8 comprises, based on 1000mL total weight, 3.5-7g of tris (hydroxymethyl) aminomethane, 7-9g of sodium chloride, 30-50g of bovine serum albumin, 80-100mL of bovine serum, 10-20mL of sheep serum, 10-20mL of horse serum, and the balance of water, wherein the pH is 5.8-7.6, and the content of biotin-IL-6 antibody is 2-4. mu.g/mL, preferably 2. mu.g/mL.
Further, the pH of the buffer is adjusted using sodium hydroxide or hydrochloric acid.
Further, the specific composition of the reagent C is a mixture of streptavidin-magnetic particles and a buffer 9;
the buffer solution 9 is specifically composed of, by total amount of 1000mL, 5.6-5.9g of disodium hydrogen phosphate dodecahydrate, 0.55-0.60g of sodium dihydrogen phosphate, 7-9g of sodium chloride, 30-50g of bovine serum albumin, 80-100g of sucrose, 3.0-5.0g of cellulose salt or cellulose derivative, 20-30g of gelatin, and the balance of water, wherein the pH value is 6.2-8.0, and the content of streptavidin-magnetic particles is 0.4-0.8 mg/mL.
Further, the pH of the buffer is adjusted using sodium hydroxide or hydrochloric acid.
Further, the kit also comprises a cleaning solution and a luminescent substrate specifically responding to alkaline phosphatase;
further, the specific composition of the cleaning solution is such that the cleaning solution contains, calculated as 1000mL of solution: 3-5g of tris (hydroxymethyl) aminomethane, 10-20g of NaCl, 3-5g of Tween 20, 3-5mL of TritonX100, and the balance of water.
Further, the alkaline phosphatase specifically responds to a luminescent substrate.
Further, the luminescent substrate specifically responding to the alkaline phosphatase is a tris solution containing a dioxetane luminescent substrate (AMPDD) or a dihydroacridine luminescent substrate.
Further, the luminescent substrate of the specific response of the alkaline phosphatase is 9- [ (p-chlorophenylthio) -phosphoryloxymethylene ] -10-methyl-9, 10-dihydroacridine disodium salt.
The term "alkaline phosphatase-IL-6 antibody" as used herein is an alkaline phosphatase-labeled IL-6 antibody known to those skilled in the art.
The term "biotin-IL-6 antibody" as used herein is intended to mean a biotin-labeled IL-6 antibody known to those skilled in the art.
The term "streptavidin-magnetic particles" as used herein is streptavidin-labeled magnetic microspheres.
The measurement unit used in the present invention is equivalent to 10pg/mL-12g/mL。
The invention has the advantages that:
1. the appropriate buffer system is adopted, so that the sensitivity and the accuracy of IL-6 detection are improved, and the detection sensitivity reaches 5 multiplied by 10-12g/mL or less.
2. The kit can be matched with a full-automatic instrument for detection, and accurate results can be obtained only by adding the blood collection tube with the sample for 18 minutes. Is far superior to the reaction time of 30 minutes of the traditional chemiluminescence or the reaction time of 1-2 hours of the enzyme-linked immunosorbent assay.
Detailed Description
Buffer solution example 1
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, and IL-6 recombinant protein, the balance being water, the total amount being 1000mL, wherein the concentration of the IL-6 recombinant protein is 15pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, thus obtaining the buffer S1-1.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, and IL-6 recombinant protein, the balance being water, the total amount being 1000mL, wherein the concentration of the IL-6 recombinant protein is 50pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, thus obtaining the buffer S1-2.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, and IL-6 recombinant protein, the balance being water, the total amount being 1000mL, wherein the concentration of the IL-6 recombinant protein is 300pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, thus obtaining the buffer S1-3.
Buffer example 2
Buffer solution suitable for interleukin-6 detection
The buffer solution S2-1 is prepared by using 18g of tris (hydroxymethyl) aminomethane, 30g of bovine serum albumin, 30g of glycine, 50g of trehalose, 30g of casein, 30g of IL-6 recombinant protein and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 10pg/mL, and the pH value is adjusted to 8.8 by using 4M HCl.
Buffer solution suitable for interleukin-6 detection
The buffer solution S2-2 is prepared by using 18g of tris (hydroxymethyl) aminomethane, 30g of bovine serum albumin, 30g of glycine, 50g of trehalose, 30g of casein, 30g of IL-6 recombinant protein and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 100pg/mL, and the pH value is adjusted to 8.8 by using 4M HCl.
Buffer solution suitable for interleukin-6 detection
The buffer solution S2-3 is prepared by the steps of preparing buffer solution S2-3, wherein the buffer solution comprises 18g of tris (hydroxymethyl) aminomethane, 30g of bovine serum albumin, 30g of glycine, 50g of trehalose, 30g of casein, 30g of IL-6 recombinant protein and the balance of water, the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 500pg/mL, and the pH value is adjusted to 8.8 by using 4M HCl.
Buffer comparative example 1-Triton X-100 was added to the buffer as compared to buffer example 1
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, 40g of Triton X-10040 g of IL-6 recombinant protein and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 15pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, so that the buffer solution D1-1 is obtained.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, 40g of Triton X-10040 g of IL-6 recombinant protein and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 50pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, so that the buffer solution D1-2 is obtained.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, 40g of Triton X-10040 g of IL-6 recombinant protein and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 300pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, so that the buffer solution D1-3 is obtained.
Buffer comparative example 2-replacement of trehalose and casein with sucrose and gelatin compared to buffer example 1
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of sucrose, 25g of gelatin, and IL-6 recombinant protein, and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 15pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, so that the buffer solution D2-1 is obtained.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of sucrose, 25g of gelatin, and IL-6 recombinant protein, and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 50pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, so that the buffer solution D2-2 is obtained.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of sucrose, 25g of gelatin, and IL-6 recombinant protein, and the balance of water, wherein the total amount is 1000mL, the concentration of the IL-6 recombinant protein is 300pg/mL, and the pH value is adjusted to 7.2 by using 4M HCl, so that the buffer solution D3-3 is obtained.
Buffer comparative example 3-higher pH compared to buffer example 1
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, and IL-6 recombinant protein, the balance being water, the total amount being 1000mL, wherein the concentration of the IL-6 recombinant protein is 15pg/mL, and the pH value is adjusted to 9.5 by using 4M HCl, thus obtaining the buffer solution D3-1.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, and IL-6 recombinant protein, the balance being water, the total amount being 1000mL, wherein the concentration of the IL-6 recombinant protein is 50pg/mL, and the pH value is adjusted to 9.5 by using 4M HCl, thus obtaining the buffer solution D3-2.
Buffer solution suitable for interleukin-6 detection
12g of tris (hydroxymethyl) aminomethane, 50g of bovine serum albumin, 20g of glycine, 40g of trehalose, 25g of casein, and IL-6 recombinant protein, the balance being water, the total amount being 1000mL, wherein the concentration of the IL-6 recombinant protein is 300pg/mL, and the pH value is adjusted to 9.5 by using 4M HCl, thus obtaining the buffer solution D3-3.
The invention also provides a preparation method of the materials required by the preparation of the kit
Buffer 1: ethanolamine solution 15g/L and NaCl solution 6 g/L;
buffer 2: 75g/L of glycine aqueous solution;
buffer 3: 203.3g/L magnesium chloride hexahydrate solution;
buffer 4: 30g/L morpholine ethanesulfonic acid sodium salt aqueous solution;
buffer 5: 5g/L of trihydroxymethyl aminomethane, 9g/L of sodium chloride and 20g/L of bovine serum albumin;
buffer 6: 6g/L of trihydroxymethyl aminomethane and 9g/L of sodium chloride.
Preparation of alkaline phosphatase-IL-6 antibody
8mg of 2-iminothiolane hydrochloride (2IT) was weighed out and dissolved in buffer 1 to 13.76 mg/mL. The 2IT solution is added into the antibody solution for activation according to the molar ratio of the 2-IT to the antibody of 30: 1. After shaking and mixing, the mixture was reacted at room temperature for 30 minutes. And stopping activation, adding the buffer solution 2 into the antibody solution according to the proportion that 1mg of the antibody is added into 5-20 mu l of the buffer solution 2, and reacting for 10min at room temperature. Excess 2IT was removed using a PD10 desalting column and the activated antibody was collected.
4mg of succinimidyl (N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) were weighed out and dissolved in Dimethylformamide (DMF) to 6.69 mg/mlL. The SMCC solution was added to the ALP solution at a 50:1 molar ratio of SMCC to ALP. After shaking and mixing, the mixture was reacted at room temperature for 30 minutes. After the termination of the activation, 1mg of ALP (alkaline phosphatase) was added to 10 to 50. mu.l of buffer 2, and buffer 2 was added to the ALP solution to react at room temperature for 10 min. Excess SMCC was removed using a PD10 desalting column and the ALP was collected after activation.
Adding ALP solution into the antibody solution according to the mass ratio of the activated antibody to the activated ALP of 1:1 (namely, adding 1.0mg of antibody into 1.0mg of ALP, shaking and mixing uniformly, and reacting the mixture for 12-18 hours in an environment of 2-8 ℃.
1-10mg of maleimide was weighed out and dissolved in DMF to 9.7 mg/mL. At a ratio of 1/10, the solution was diluted with buffer 1 to give a 0.97mg/mL maleimide solution. This solution was added in a ratio of 1mg of the antibody to 10. mu.l of a 0.97mg/mL maleimide solution, and reacted at room temperature for 15 minutes. mu.L of ethanolamine was measured accurately and dissolved in buffer 1 to 100 mM. That is, 994. mu.L of buffer 1 was added to 6. mu.L of ethanolamine. Adding 10-50 mul of 100mM ethanolamine solution into 1mg of antibody, and shaking and mixing uniformly. And concentrating the antibody conjugate to be purified to 0.5-2 mg/mL by using an ultrafiltration concentration tube. Antibody purification was performed using a purified protein analyzer and Superdex 200 preparative 2.6/60 gel column, buffer 2 as eluent. The purified liquid is alkaline phosphatase-IL-6 antibody solution.
Preparation example of biotin-IL-6 antibody
A10 mM biotin solution was prepared in 0.02M PBS. 10mM biotin solution was added to the antibody solution at a molar ratio of 1:20 antibody to biotin. After shaking and mixing, the mixture was reacted at room temperature for 1 hour.
The reacted solution was added to a desalting column. When the solution completely entered the column, the column volume was made up with 0.02M PBS. When the liquid in the column completely enters the column material, an equal volume of 0.02M PBS buffer is added to start elution. Equal volumes of protein eluate were collected and tested for concentration. The eluted solution is the biotin-IL-6 antibody solution.
Preparation example of streptavidin-magnetic microparticle
After washing the carboxyl group-containing magnetic microparticles with buffer 4, they were resuspended at 10 mg/mL. According to the mass ratio of the magnetic particles to the streptavidin of 10: 1 streptavidin was added to the magnetic particle solution. After thorough mixing, the reaction mixture was mixed well at room temperature for 10 minutes. EDC is added to the magnetic particle solution in a ratio of the magnetic particles to (3-dimethylaminopropyl) ethylcarbodiimide hydrochloride (EDC) mass ratio of 100: 10. After thorough mixing, the reaction mixture was mixed at room temperature for 2 hours. Then, the magnetic separation is carried out on the connecting object, the separated magnetic particles are resuspended to 10mg/mL by using buffer solution 6, and the magnetic particles are preserved in the environment of 2-8 ℃.
Kit example 1 IL-6 detection kit
Reagent A: specifically, the alkaline phosphatase-IL-6 antibody was composed of 5g of tris, 9g of sodium chloride, 50g of bovine serum albumin, 100mL of bovine serum, 15mL of sheep serum, 15mL of horse serum, and the balance of water, and the pH was adjusted to 7.6 using 4M HCl, wherein the content of the alkaline phosphatase-IL-6 antibody was 2 μ g/mL.
And (3) reagent B: specifically, the biotin-IL-6 antibody was composed of, based on a total amount of 1000mL, 5g of tris, 9g of sodium chloride, 50g of bovine serum albumin, 100mL of bovine serum, 15mL of sheep serum, 15mL of horse serum, and the balance of water, and the pH was adjusted to 7.6 using 4M HCl, wherein the content of the biotin-IL-6 antibody was 2 μ g/mL.
And (3) reagent C: the streptavidin-magnetic particles are specifically composed of, based on a total volume of 1000mL, 5.9g of disodium hydrogen phosphate dodecahydrate, 0.6g of sodium dihydrogen phosphate, 9g of sodium chloride, 50g of bovine serum albumin, 100g of sucrose, 5.0g of cellulose, 30g of gelatin, and the balance of water, and the pH is adjusted to 8.0 using 4M HCl, wherein the content of the streptavidin-magnetic particles is 0.4 mg/mL.
The cleaning solution comprises, by 1000mL, 3-5g of tris (hydroxymethyl) aminomethane, 10-20g of NaCl, 3-5g of Tween 20, 3-5mL of TritonX-100, and the balance of water.
A luminescent substrate specifically responsive to alkaline phosphatase, 9- [ (p-chlorophenylthio) -phosphoryloxymethylene ] -10-methyl-9, 10-dihydroacridine disodium salt, in tris (hydroxymethyl) aminomethane at a concentration of 0.3 mg/mL.
Buffer S1-1, buffer S1-2 and buffer S1-3
Kit embodiment 2 IL-6 detection kit
Reagent A: specifically, the alkaline phosphatase-IL-6 antibody was composed of 5g of tris, 9g of sodium chloride, 50g of bovine serum albumin, 100mL of bovine serum, 15mL of sheep serum, 15mL of horse serum, and the balance of water, and the pH was adjusted to 7.6 using 4M HCl, wherein the content of the alkaline phosphatase-IL-6 antibody was 2 μ g/mL.
And (3) reagent B: specifically, the biotin-IL-6 antibody was composed of, based on a total amount of 1000mL, 5g of tris, 9g of sodium chloride, 50g of bovine serum albumin, 100mL of bovine serum, 15mL of sheep serum, 15mL of horse serum, and the balance of water, and the pH was adjusted to 7.6 using 4M HCl, wherein the content of the biotin-IL-6 antibody was 2 μ g/mL.
And (3) reagent C: the streptavidin-magnetic particles are specifically composed of, based on a total volume of 1000mL, 5.9g of disodium hydrogen phosphate dodecahydrate, 0.6g of sodium dihydrogen phosphate, 9g of sodium chloride, 50g of bovine serum albumin, 100g of sucrose, 5.0g of cellulose, 30g of gelatin, and the balance of water, and the pH is adjusted to 8.0 using 4M HCl, wherein the content of the streptavidin-magnetic particles is 0.4 mg/mL.
Cleaning fluid;
the cleaning solution comprises, by 1000mL, 3-5g of tris (hydroxymethyl) aminomethane, 10-20g of NaCl, 3-5g of Tween 20, 3-5mL of TritonX100, and the balance of water.
A luminescent substrate that specifically responds to alkaline phosphatase; a solution of 9- [ (p-chlorophenylthio) -phosphoryloxymethylene ] -10-methyl-9, 10-dihydroacridine disodium salt in tris (hydroxymethyl) aminomethane at a concentration of 0.3 mg/mL.
Buffer S2-1, buffer S2-2 and buffer S2-3
Comparative kit example 1
The difference with respect to kit example 1 is that the buffer liquid system is:
buffer D1-1, buffer D1-2 and buffer D1-3.
Comparative kit example 2
The difference with respect to kit example 1 is that the buffer liquid system is:
buffer D2-1, buffer D2-2 and buffer D2-3.
Comparative kit example 3
The difference with respect to kit example 1 is that the buffer liquid system is:
buffer D3-1, buffer D3-2 and buffer D3-3.
Performance test index of reagent kit
The test method comprises the following steps:
immune reaction: mu.L of the sample, 50. mu.L of reagent A, 50. mu.L of reagent B, and 50. mu.L of reagent C were mixed in this order in a centrifuge tube 1, and reacted at 37 ℃ for 20 min.
Magnetic separation and cleaning: after adding 300. mu.L of the washing solution and washing for 2min, the mixture containing the magnetic particles is magnetically sucked out of the centrifuge tube 1 and demagnetized in the centrifuge tube 2. Repeat 3 times.
Reading value: after adding 150. mu.L of luminescent substrate to the washed magnetic particles, the luminescent substrate catalyzed by ALP emits light and then the relative luminescence intensity (RLU) is measured by a self-developed instrument.
And obtaining an IL-6 concentration-luminous value standard curve according to the detected value of the calibrator. The curve was fitted using a four parameter Logistic equation.
The detection value of the sample can correspond to the unique concentration value obtained on the curve, so that the concentration detection of the unknown sample is realized.
The accuracy analysis method comprises the following steps:
interleukin 6(IL-6) solution (A) was added to a sample B at a concentration of about 300pg/mL (tolerance. + -. 10%) in a volume ratio of 1:9 between the IL-6 antigen added and the sample B, and the recovery rate R was calculated according to equation (1) and should be in the range of 85% to 115%.
In the formula:
r-recovery rate;
v is the volume of the sample A liquid;
V0-volume of serum sample B fluid;
c is the average value of 3 measurements after the serum sample B liquid is added into the liquid A;
C0-mean of 3 measurements of serum sample B fluid;
CS-concentration of sample a liquid.
Blank limit analysis method:
repeating the test for 20 times to obtain concentration values of 20 test results, and calculating average value
And Standard Deviation (SD). Mean value of
The blank limit is obtained, and the result is less than or equal to 5 pg/mL.
Linear interval analysis method:
mixing a high value sample close to the upper limit of the linear region and a low value sample close to the lower limit of the linear region or a zero concentration sample to obtain not less than 5 dilution concentrations, wherein the low value concentration sample is close to the lower limit of the linear region. The test was repeated 3 times for each concentration of the sample to obtain the luminescence value, the measurement result of each sample was recorded, and the average value (y) of the 3 measurements of each sample was calculatedi). In diluted concentration (x)i) As independent variable, the mean value (y) of the results is determinedi) Linear regression equations were solved for the dependent variables. And (3) calculating a correlation coefficient (r) of the linear regression according to the formula (2), and determining a linear interval of which the correlation coefficient r is more than or equal to 0.990.
In the formula:
r- — -correlation coefficient;
xi-dilution ratio;
yi-mean value of individual sample measurements;
Method for analyzing repeatability
The quality control product is tested repeatedly for 10 times by the same batch number kit, and the average value of 10 test results is calculated
And standard deviation SD. The Coefficient of Variation (CV) was calculated according to equation (3) and the result CV was less than or equal to 8%.
In the formula: s-standard deviation of sample test values;
The method for analyzing the difference between batches comprises the following steps:
the quality control materials are tested repeatedly for 10 times by using the kits with 3 batch numbers respectively, and the average value of the test results of 30 times is calculated
And standard deviation SD, and obtaining Coefficient of Variation (CV) according to formula (3), wherein the result CV is less than or equal to 12%.
The test was performed using the above method for kit examples 1-2 and kit comparative examples 1-3, and the results are shown in Table 1.
TABLE 1 test results
| Accuracy of | Margin limit | Difference between batches | Coefficient of correlation (r) | Repeatability (CV) | |
| Kit example 1 | 102.51％ | 3.305 | 8.63％ | 0.9987 | 6.15％ |
| Kit example 2 | 97.26％ | 4.231 | 10.27％ | 0.9992 | 6.33％ |
| Comparative kit example 1 | 82.33％ | 7.328 | 16.92％ | 0.9885 | 9.47％ |
| Comparative kit example 2 | 131.46％ | 6.929 | 29.69％ | 0.8612 | 12.42％ |
| Comparative kit example 3 | 76.37％ | 5.326 | 19.36％ | 0.8818 | 10.58％ |
It can be seen that the kit composed of the buffer solution of the invention has a better lower margin limit, a larger linear interval and linear relation in IL-6 detection, and better repeatability.
Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, and that the simple modifications or equivalent substitutions of the technical solutions of the present invention by those of ordinary skill in the art can be made without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A buffer solution suitable for detecting interleukin-6 comprises the following components,
12-18 parts of trihydroxymethyl aminomethane, 30-50 parts of bovine serum albumin, 20-30 parts of glycine, 30-50 parts of trehalose and 20-30 parts of casein.
2. The buffer according to claim 1, comprising the following components in a volume of 1000mL,
12-18g of tris (hydroxymethyl) aminomethane, 30-50g of bovine serum albumin, 20-30g of glycine, 30-50g of trehalose, 20-30g of casein, IL-6 recombinant protein and the balance of water, wherein the concentration of the IL-6 recombinant protein is 10-500 pg/mL.
3. A buffer according to claim 1 or 2, wherein the pH is 7.2 to 8.8.
4. Use of a buffer according to any of claims 1-3 in the preparation of an IL-6 assay product.
5. A kit comprising the buffer of any of claims 2-3, comprising the following components:
reagent A: alkaline phosphatase-IL-6 antibody;
and (3) reagent B: biotin-IL-6 antibodies;
and (3) reagent C: streptavidin-magnetic microparticles;
and three different concentrations of the buffer according to any one of claims 2 to 3, the three different concentrations of the buffer according to any one of claims 2 to 3 differing only in the concentration of the recombinant IL-6 protein.
6. The kit as claimed in claim 5, wherein the concentrations of IL-6 in the three buffers with different concentrations are 110-15 pg/mL standard, 2290-310 pg/mL standard and 40-50pg/mL quality control.
7. The kit according to claim 5, wherein the specific composition of the reagent A is a mixture of alkaline phosphatase-IL-6 antibody and buffer 8;
the buffer solution 8 comprises, based on 1000mL total weight, 3.5-7g of tris (hydroxymethyl) aminomethane, 7-9g of sodium chloride, 30-50g of bovine serum albumin, 80-100mL of bovine serum, 10-20mL of sheep serum, 10-20mL of horse serum, and the balance of water, wherein the pH value is 5.8-7.6, and the content of alkaline phosphatase-IL-6 antibody is 2-4. mu.g/mL, preferably 2. mu.g/mL.
8. The kit according to claim 5, wherein the specific composition of reagent B is a mixture of biotin-IL-6 antibody and buffer 8;
the buffer solution 8 comprises, based on 1000mL total weight, 3.5-7g of tris (hydroxymethyl) aminomethane, 7-9g of sodium chloride, 30-50g of bovine serum albumin, 80-100mL of bovine serum, 10-20mL of sheep serum, 10-20mL of horse serum, and the balance of water, wherein the pH is 5.8-7.6, and the content of biotin-IL-6 antibody is 2-4. mu.g/mL, preferably 2. mu.g/mL.
9. The kit according to claim 5, wherein the reagent C is composed of a mixture of streptavidin-magnetic particles and buffer 9;
the buffer solution 9 is specifically composed of, by total amount of 1000mL, 5.6-5.9g of disodium hydrogen phosphate dodecahydrate, 0.55-0.60g of sodium dihydrogen phosphate, 7-9g of sodium chloride, 30-50g of bovine serum albumin, 80-100g of sucrose, 3.0-5.0g of cellulose salt or cellulose derivative, 20-30g of gelatin, and the balance of water, wherein the pH value is 6.2-8.0, and the content of streptavidin-magnetic particles is 0.4-0.8 mg/mL.
10. The kit of claim 5, further comprising a wash solution and a luminescent substrate that is specifically responsive to alkaline phosphatase;
the specific composition of the cleaning solution is calculated by 1000mL of solution, and the cleaning solution contains: 3-5g of tris (hydroxymethyl) aminomethane, 10-20g of NaCl, 3-5g of Tween 20, 3-5mL of TritonX100, and the balance of water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110465801.5A CN113203863B (en) | 2021-04-28 | 2021-04-28 | Buffer solution suitable for interleukin-6 detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110465801.5A CN113203863B (en) | 2021-04-28 | 2021-04-28 | Buffer solution suitable for interleukin-6 detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113203863A true CN113203863A (en) | 2021-08-03 |
| CN113203863B CN113203863B (en) | 2022-01-21 |
Family
ID=77029191
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110465801.5A Active CN113203863B (en) | 2021-04-28 | 2021-04-28 | Buffer solution suitable for interleukin-6 detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113203863B (en) |
Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006029652A1 (en) * | 2004-09-15 | 2006-03-23 | Medizinische Hochschule Hannover | Use of ilt6 for immunomodulation |
| CN103261222A (en) * | 2010-09-10 | 2013-08-21 | 埃洛齐纳公司 | Novel antibody derivatives |
| CN104330551A (en) * | 2014-11-17 | 2015-02-04 | 南方医科大学南方医院 | Interleukin 6 quantitative determination kit and preparation method thereof |
| CN105277717A (en) * | 2015-09-28 | 2016-01-27 | 成都博奥新景医学科技有限公司 | Magnetic particle separation and chemiluminescence immunoassay method of thyroglobulin |
| CN105445466A (en) * | 2016-01-19 | 2016-03-30 | 苏州市博纳泰科生物技术有限公司 | Detection method for interleukin 6 and reagent kit of detection method |
| CN107402308A (en) * | 2017-06-21 | 2017-11-28 | 北京科跃中楷生物技术有限公司 | Immue quantitative detection reagent boxes of IL 6 and preparation method thereof |
| CN108037285A (en) * | 2017-11-29 | 2018-05-15 | 北京美联泰科生物技术有限公司 | A kind of magnetic microparticle chemiluminescence quantitatively detects kit of UCHL-1 and preparation method thereof |
| CN108398557A (en) * | 2018-02-09 | 2018-08-14 | 河南省生物工程技术研究中心有限公司 | A kind of myeloperoxidase fluorescence immune chromatography test paper bar and test card |
| CN108409852A (en) * | 2018-02-07 | 2018-08-17 | 深圳市新产业生物医学工程股份有限公司 | Sex hormone binding globulin preserves liquid and its application |
| CN108722507A (en) * | 2018-06-21 | 2018-11-02 | 微粒云科技（北京）有限公司 | A kind of inflammation four marker magnetic particle microflow controlled biochips, detection methods |
| CN109917134A (en) * | 2018-12-21 | 2019-06-21 | 广州市进德生物科技有限公司 | A kind of calibration object stabilizer, the detection kit and detection method for measuring C peptide |
| CN110261621A (en) * | 2019-07-16 | 2019-09-20 | 三诺生物传感股份有限公司 | A kind of interleukin-6 detection kit |
| CN110988367A (en) * | 2019-11-18 | 2020-04-10 | 迈克生物股份有限公司 | Storage agent, calibrator for detecting LH and detection kit |
| CN111505303A (en) * | 2019-01-31 | 2020-08-07 | 艾维可生物科技有限公司 | Kit for detecting heart-type fatty acid binding protein by chemiluminescence method and use method thereof |
| CN112129950A (en) * | 2020-09-11 | 2020-12-25 | 武汉生之源生物科技股份有限公司 | Magnetic particle chemiluminescence kit for detecting interleukin 6 |
| CN112630430A (en) * | 2020-11-16 | 2021-04-09 | 北京美联泰科生物技术有限公司 | Kit for quantitatively detecting UCHL-1 and application thereof |
-
2021
- 2021-04-28 CN CN202110465801.5A patent/CN113203863B/en active Active
Patent Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006029652A1 (en) * | 2004-09-15 | 2006-03-23 | Medizinische Hochschule Hannover | Use of ilt6 for immunomodulation |
| CN103261222A (en) * | 2010-09-10 | 2013-08-21 | 埃洛齐纳公司 | Novel antibody derivatives |
| CN104330551A (en) * | 2014-11-17 | 2015-02-04 | 南方医科大学南方医院 | Interleukin 6 quantitative determination kit and preparation method thereof |
| CN105277717A (en) * | 2015-09-28 | 2016-01-27 | 成都博奥新景医学科技有限公司 | Magnetic particle separation and chemiluminescence immunoassay method of thyroglobulin |
| CN105445466A (en) * | 2016-01-19 | 2016-03-30 | 苏州市博纳泰科生物技术有限公司 | Detection method for interleukin 6 and reagent kit of detection method |
| CN107402308A (en) * | 2017-06-21 | 2017-11-28 | 北京科跃中楷生物技术有限公司 | Immue quantitative detection reagent boxes of IL 6 and preparation method thereof |
| CN108037285A (en) * | 2017-11-29 | 2018-05-15 | 北京美联泰科生物技术有限公司 | A kind of magnetic microparticle chemiluminescence quantitatively detects kit of UCHL-1 and preparation method thereof |
| CN108409852A (en) * | 2018-02-07 | 2018-08-17 | 深圳市新产业生物医学工程股份有限公司 | Sex hormone binding globulin preserves liquid and its application |
| CN108398557A (en) * | 2018-02-09 | 2018-08-14 | 河南省生物工程技术研究中心有限公司 | A kind of myeloperoxidase fluorescence immune chromatography test paper bar and test card |
| CN108722507A (en) * | 2018-06-21 | 2018-11-02 | 微粒云科技（北京）有限公司 | A kind of inflammation four marker magnetic particle microflow controlled biochips, detection methods |
| CN109917134A (en) * | 2018-12-21 | 2019-06-21 | 广州市进德生物科技有限公司 | A kind of calibration object stabilizer, the detection kit and detection method for measuring C peptide |
| CN111505303A (en) * | 2019-01-31 | 2020-08-07 | 艾维可生物科技有限公司 | Kit for detecting heart-type fatty acid binding protein by chemiluminescence method and use method thereof |
| CN110261621A (en) * | 2019-07-16 | 2019-09-20 | 三诺生物传感股份有限公司 | A kind of interleukin-6 detection kit |
| CN110988367A (en) * | 2019-11-18 | 2020-04-10 | 迈克生物股份有限公司 | Storage agent, calibrator for detecting LH and detection kit |
| CN112129950A (en) * | 2020-09-11 | 2020-12-25 | 武汉生之源生物科技股份有限公司 | Magnetic particle chemiluminescence kit for detecting interleukin 6 |
| CN112630430A (en) * | 2020-11-16 | 2021-04-09 | 北京美联泰科生物技术有限公司 | Kit for quantitatively detecting UCHL-1 and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| WU D 等: "Association of peripheral cytokine levels with cerebral structural abnormalities in schizophrenia", 《BRAIN RES》 * |
| 孙飞 等: "IMMUNITE1000化学发光分析系统检测白介素-6的性能验证", 《中国中西医结合学会.第一次全国中西医结合检验医学学术会议暨中国中西医结合学会检验医学专业委员会成立大会论文汇编[C].中国中西医结合学会:中国中西医结合学会》 * |
| 朱里等: "SLE患者HSP90基因表达及其与IL-6相关性的研究", 《中国麻风皮肤病杂志》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113203863B (en) | 2022-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108008132B (en) | Kit for combined detection of ovarian cancer tumor markers HE4 and CA125 and preparation method and application thereof | |
| CN112630430B (en) | Kit for quantitatively detecting UCHL-1 and application thereof | |
| CN113804896A (en) | Magnetic particle chemiluminescence kit for measuring interleukin-8 content in human serum | |
| CN111089958A (en) | P16 based on glucan signal amplificationINK4aChemiluminescence kit | |
| CN202916286U (en) | Latex enhanced turbidimetric immunoassay kit for quantitatively detecting procalcitonin (PCT) | |
| CN113614539A (en) | Quantitative kit for myxovirus resistance protein 1 | |
| CN112269025A (en) | Interleukin-6 chemiluminescence assay kit and preparation method thereof | |
| CN113203867B (en) | Buffer solution suitable for insulin detection kit | |
| CN113203863B (en) | Buffer solution suitable for interleukin-6 detection | |
| CN113933521A (en) | Magnetic particle chemiluminescence detection kit and preparation method and application thereof | |
| CN106366199B (en) | Troponin I monoclonal antibody magnetic particle, preparation method thereof and detection kit | |
| CN113203868B (en) | Buffer solution suitable for AMH detection kit | |
| CN108362892B (en) | Procalcitonin colloidal gold immunoturbidimetry detection reagent | |
| CN111273033A (en) | Golgi protein73 determination kit and chemiluminescence determination method thereof | |
| CN110988330A (en) | Mycobacterium tuberculosis specific cellular immune reaction magnetic particle chemiluminescence detection kit and preparation method thereof | |
| CN115353566A (en) | Antibody combination for detecting interleukin 1-beta and application thereof | |
| CN113125761A (en) | Composition for procalcitonin detection, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method | |
| CN114216897A (en) | sST2 chemiluminescence detection kit and detection method thereof | |
| US6927071B2 (en) | Method for reducing non-specific aggregation of latex microparticles in the presence of serum or plasma | |
| CN111044734A (en) | Detection kit for detecting vascular endothelial growth factor, and preparation method and use method thereof | |
| CN117405890B (en) | Chemiluminescent detection kit and method for pepsinogen II | |
| WO2023096393A1 (en) | Composition for improving diagnostic performance of immunoassay comprising avidin-expressing microorganisms | |
| CN111381046A (en) | Calprotectin chemiluminescence immunoassay kit and preparation method thereof | |
| CN113092763B (en) | Buffer solution and application thereof in neuron specific enolase detection kit | |
| CN111721937B (en) | Human epididymis protein 4 immunoturbidimetry kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |