CN114112957A - Adiponectin determination kit and application thereof - Google Patents
Adiponectin determination kit and application thereof Download PDFInfo
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Abstract
The invention provides an adiponectin determination kit and application thereof, and relates to the technical field of biological detection. According to the invention, a stabilizing agent and a surfactant are added into a reagent R1, the stabilizing agent is mannitol and chitosan with the mass ratio of 1-5:1, the surfactant is a mixture of sophorolipid, sodium dodecyl benzene sulfonate and polyethylene glycol 6000 with the mass ratio of 1:1-3:5, and the specific components and concentration ratio of the sophorolipid, the sodium dodecyl benzene sulfonate and the polyethylene glycol 6000 are controlled, so that the sensitivity and the accuracy of the kit are obviously improved; by controlling the mass ratio of sophorolipid to chitosan, the stability of the kit can be obviously improved, and the kit still has good stability after being placed in a dark and sealed condition at the temperature of 2-8 ℃ for 16 months even under the condition that a preservative is not added into the reagent R1.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to an adiponectin determination kit and application thereof.
Background
Adiponectin is an adipocyte factor that is stable in human plasma at concentrations ranging from about 3.0 to about 30.0mg/L and represents about 0.01% of plasma protein. Human adiponectin is 244 amino acids and includes 3 regions: an amino-terminal signal sequence, a collagen domain, and a carboxy-terminal globular domain. Adiponectin exists in blood as three subtypes, trimer (65kb), hexamer (150kb), and high molecular weight polymer (18-36 multimers, >280 kb). Clinical studies show that adiponectin is closely related to obesity, type II diabetes, coronary heart disease, and insulin resistance, and has anti-atherogenesis, anti-inflammation, and anti-intimal hyperplasia properties after vascular injury.
The adiponectin can be measured by various methods, such as Radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA), for example, Chinese patent application 200910043672X discloses a human adiponectin ELISA kit, a preparation method and an application thereof, wherein the kit comprises an enzyme label plate coated by an anti-human APN monoclonal antibody, a recombinant gAPN protein standard substance, an anti-human APN monoclonal detection antibody, HRP-streptomycin, 10mM phosphate buffer solution with pH7.4, TMB enzyme substrate color development liquid, 0.1M phosphate buffer solution with pH7.4 and 2M H2SO4. The invention also comprises a preparation method of the kit and application of the kit in detection of APN content in human blood. Although the ELISA method is clinically used for nearly twenty years, it still has some fatal disadvantages, long operation time and low automation degree. Radioimmunoassay (RIA) is highly sensitive, but is unstable, less reproducible than ELISA, and poses the risk of radioactive contamination.
The latex enhanced immunoturbidimetry is currently used more frequently, and has the following advantages: saving time and labor: the antigen and antibody reaction is carried out in a homogeneous reaction system, and the result can be obtained in several minutes by directly measuring the absorbance value of the reaction solution by using a full-automatic biochemical analyzer, so that the complicated operation steps of repeated incubation and plate washing of an ELISA method and the like are omitted. Secondly, the method is stable and accurate: the simplification of the operation steps correspondingly avoids the interference of a plurality of human operation factors and external factors such as reagents, environment and the like, has better stability and repeatability, and can reflect the content of the adiponectin in the tested human serum more truly. The application is wide: the immunoturbidimetry is less sensitive than ELISA, but has a lower limit sufficient for detecting adiponectin in human serum, and can completely meet the clinical detection requirements.
For example, chinese patent application 202010101938.8 discloses a latex-enhanced immunoturbidimetric assay kit for quantitatively detecting adiponectin ADPN and a preparation and application method thereof, comprising an adiponectin R1 reagent and an adiponectin R2 reagent; the adiponectin R1 reagent comprises a buffer solution A, a protective agent A, a reaction enhancer and a preservative; the adiponectin R2 reagent comprises a buffer solution B, a protective agent B, a preservative and sensitized polystyrene latex particles coated with an anti-human adiponectin antibody; wherein the anti-human adiponectin antibodies in the sensitized polystyrene latex particles coated with the anti-human adiponectin antibodies are connected with the sensitized polystyrene latex particles through a streptavidin-biotin system. The invention also provides a preparation method and an application method of the latex enhanced immunoturbidimetric kit for quantitatively detecting adiponectin ADPN. However, the stability, reproducibility and accuracy of the given kit are not satisfactory.
Therefore, it is required to provide an adiponectin measurement kit with good stability and high repeatability and accuracy and a preparation method thereof.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the adiponectin determination kit with good stability and high repeatability and accuracy and the preparation method thereof.
The invention adopts the following technical scheme to solve the technical problems:
an adiponectin assay kit comprising a R1 reagent and a R2 reagent.
The R1 reagent comprises a buffer solution, an electrolyte, a stabilizer and a surfactant A;
the R2 reagent comprises anti-human adiponectin antibody latex particles, buffer solution, surfactant B and preservative.
The surfactant A is one or more of sophorolipid, sodium dodecyl benzene sulfonate, polyethylene glycol mono-octyl phenyl ether, polyethylene glycol 6000, triton X-100 and polyvinylpyrrolidone.
Preferably, the surfactant is one or more of sophorolipid, sodium dodecyl benzene sulfonate, polyethylene glycol mono-octyl phenyl ether and polyethylene glycol 6000;
more preferably, the surfactant is a mixture of sophorolipid, sodium dodecylbenzenesulfonate and polyethylene glycol 6000;
further preferably, the surfactant is a mixture of sophorolipid, sodium dodecyl benzene sulfonate and polyethylene glycol 6000 in a mass ratio of 1:1-3: 5;
still further preferably, the surfactant is a mixture of sophorolipid, sodium dodecylbenzenesulfonate and polyethylene glycol 6000 in a mass ratio of 1:2: 5.
The surfactant B is one or more of Tween-20, Tween-80, span-60, span-80 and Triton X-100;
preferably, the surfactant B is Tween-80.
The stabilizer is one or more of mannitol, sorbitol, trehalose, sucrose and chitosan;
preferably, the stabilizing agent is mannitol and chitosan in a mass ratio of 1-5: 1;
still preferably, the stabilizer is mannitol and chitosan in a mass ratio of 2-4: 1;
further preferably, the stabilizer is mannitol and chitosan in a mass ratio of 3: 1.
The buffer solution is one or more of phosphate buffer solution, acetate buffer solution, citric acid buffer solution and Tris buffer solution, and is preferably Tris buffer solution.
The electrolyte is one or more of sodium chloride, potassium chloride, magnesium chloride and magnesium sulfate, and preferably sodium chloride.
The preservative is selected from one of sodium azide, Proclin-950 or Proclin-300, and is preferably Proclin-300.
In some preferred embodiments, the reagent R1 further comprises bovine serum albumin.
In some preferred embodiments, the concentration ratio of sophorolipid to chitosan is 5-15: 1; preferably, the concentration ratio of the sophorolipid to the chitosan is 8-12: 1; still preferably, the concentration ratio of sophorolipid to chitosan is 12: 1.
In the implementation process, the invention unexpectedly discovers that the stability of the kit can be obviously improved by controlling the mass ratio of the sophorolipid to the chitosan, and the kit still has good stability after being placed for 16 months for a long time even under the condition that a preservative is not added into the reagent R1.
The adiponectin determination kit is characterized in that the R1 reagent comprises 90-120mmol/L of buffer solution, 3-8g/L of electrolyte, 2-5g/L of mannitol, 1-3g/L of chitosan, 10-15g/L of sophorolipid, 20-40g/L of sodium dodecyl benzene sulfonate and 600050-80 g/L of polyethylene glycol;
the R2 reagent comprises 8-12mg/L of anti-human adiponectin antibody latex particles, 90-120mmol/L of buffer solution, 100g/L of Tween-8060 and 0.1-0.5g/L of preservative.
Preferably, the adiponectin determination kit, the R1 reagent comprises 90-120mmol/L of trihydroxymethyl aminomethane with pH of 7, 3-8g/L of sodium chloride, 2-5g/L of mannitol, 1-3g/L of chitosan, 10-15g/L of sophorolipid, 20-40g/L of sodium dodecyl benzene sulfonate and 600050-80 g/L of polyethylene glycol;
the R2 reagent comprises 90-120mmol/L of trihydroxymethyl aminomethane of anti-human adiponectin antibody latex particles of 8-12mg/L, pH ═ 7, 100g/L of Tween-8060-.
Still preferably, in the adiponectin measurement kit, the R1 reagent comprises tris (hydroxymethyl) aminomethane 100mmol/L at pH7, sodium chloride 5g/L, mannitol 3g/L, chitosan 1g/L, sophorolipid 12g/L, sodium dodecylbenzenesulfonate 24g/L, and polyethylene glycol 600060 g/L;
the R2 reagent comprises anti-human adiponectin antibody latex particles of 10mg/L, pH ═ 7 of tris (hydroxymethyl) aminomethane 100mmol/L, Tween-8080 g/L and Proclin-3000.2 g/L.
The adiponectin determination kit further comprises an adiponectin quality control product and an adiponectin calibrator.
The invention also provides application of the adiponectin determination kit in quantitative determination of adiponectin content.
The invention also provides a using method of the adiponectin determination kit.
The kit is suitable for full-automatic biochemical analyzers such AS Hitachi 7170/7600/7180/008AS/3500, Meyer BS2000, Dirui CS600, Yapeh C16000, Toshiba TBA120FR/TBA-FX8, Beckman AU5800/AU680/AU2700/DXC800, Siemens ADVIA2400/CH930, Roche C701/C702, DuPont AR and the like.
The determination method of the determination kit is an end-point method, and the reaction direction is liter reaction:
(1) uniformly mixing 150 mu L of reagent R1 and 2 mu L of sample to be detected, incubating for 5min at 37 ℃ to obtain reaction liquid 1, determining absorbance A1, and determining dominant wavelength as 570 nm;
(2) adding 50 μ L reagent R2, mixing, reacting for 5min to obtain reaction solution 2, measuring absorbance A2, and measuring dominant wavelength at 570 nm;
(3) according to the steps (1) to (2), measuring the absorbance of the adiponectin standard substance with different concentrations to obtain an absorbance A blank and an absorbance A standard;
(4) drawing a standard curve by taking the absorbance change value delta A (A standard-A blank) of the standard substance as a vertical coordinate and the corresponding concentration c of the standard substance as a horizontal coordinate to obtain a standard curve equation;
(5) and (4) calculating the absorbance change value delta A (A2-A1) of the sample, and obtaining the concentration of the adiponectin in the sample according to a standard curve equation.
Compared with the prior art, the invention has the beneficial effects that:
(1) according to the adiponectin determination kit provided by the invention, the stabilizer and the surfactant are added into the reagent R1, the stabilizer is controlled to be mannitol and chitosan in a mass ratio of 1-5:1, the surfactant is a mixture of sophorolipid, sodium dodecyl benzene sulfonate and polyethylene glycol 6000 in a mass ratio of 1:1-3:5, and the specific components and concentration ratio of the sophorolipid, the sodium dodecyl benzene sulfonate and the polyethylene glycol 6000 are controlled, so that the sensitivity and the accuracy of the kit are remarkably increased;
(2) in the implementation process, the invention unexpectedly discovers that the stability of the kit can be obviously improved by controlling the mass ratio of the sophorolipid to the chitosan, and the kit still has good stability after being placed in a dark and sealed manner at the temperature of 2-8 ℃ for 16 months even under the condition that a preservative is not added to the reagent R1;
(3) the adiponectin determination kit provided by the invention can obviously improve the anti-interference capability of the kit by controlling the specific components and concentration ratio of each component in the reagent R1 and the reagent R2, wherein hemoglobin in a sample is less than or equal to 5.0g/L, bilirubin is less than or equal to 500mg/L, ascorbic acid is less than or equal to 10.0g/L, and triglyceride is less than or equal to 10.0g/L, so that the test result is not obviously interfered;
(4) the adiponectin determination kit provided by the invention has the advantages of simple and easily obtained components and low manufacturing cost.
Drawings
FIG. 1 is a standard curve of adiponectin measurement kit
FIG. 2 is a linear range curve of the adiponectin measurement kit prepared in example 1 of the present invention;
FIG. 3 is a linear range curve of the adiponectin measurement kit prepared in example 2 of the present invention;
FIG. 4 is a linear range curve of the adiponectin measurement kit prepared in example 3 of the present invention;
FIG. 5 Linear Range Curve of adiponectin assay kit prepared in example 4 of the present invention.
Detailed Description
The present invention will be further explained with reference to specific embodiments in order to make the technical means, the original characteristics, the achieved objects and the effects of the present invention easy to understand, but the following embodiments are only preferred embodiments of the present invention, and not all embodiments are possible. Based on the embodiments in the implementation, other embodiments obtained by those skilled in the art without any creative efforts belong to the protection scope of the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified, and materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Test conditions and methods:
the instrument comprises the following steps: hitachi 7170 full-automatic biochemical analyzer
Parameters are as follows:
| dominant wavelength | 570nm | Sample (I) | 2μL |
| Sub-wavelength | / | Reagent 1 | 150μL |
| Reaction temperature | 37℃ | Reagent 2 | 50μL |
| Reaction direction | Liter reaction | Type of reaction | End point method |
The method comprises the following operation steps:
EXAMPLE 1 adiponectin measurement kit
The components and the concentrations thereof are as follows:
the R1 reagent includes: 90mmol/L of trihydroxymethyl aminomethane with pH value of 7, 3g/L of sodium chloride, 2g/L of mannitol, 1g/L of chitosan, 10g/L of sophorolipid, 20g/L of sodium dodecyl benzene sulfonate and 600050 g/L of polyethylene glycol;
the R2 reagent includes: 90mmol/L of trihydroxymethyl aminomethane, 8mg/L, pH ═ 7 in latex particles of anti-human adiponectin antibody, 80g/L of Tween-8060 g/L and 3000.1 g/L of Proclin.
Example 2 adiponectin measurement kit
The components and the concentrations thereof are as follows:
the R1 reagent includes: 120mmol/L of trihydroxymethyl aminomethane with pH of 7, 8g/L of sodium chloride, 5g/L of mannitol, 3g/L of chitosan, 15g/L of sophorolipid, 40g/L of sodium dodecyl benzene sulfonate and 600080 g/L of polyethylene glycol;
the R2 reagent includes: 120mmol/L of trihydroxymethyl aminomethane, 12mg/L, pH ═ 7 of latex particles of anti-human adiponectin antibody, 80100 g/L of tween-and 3000.5 g/L of Proclin.
EXAMPLE 3 adiponectin measurement kit
The components and the concentrations thereof are as follows:
the R1 reagent includes: 110mmol/L of trihydroxymethyl aminomethane with pH of 7, 6g/L of sodium chloride, 4g/L of mannitol, 2g/L of chitosan, 13g/L of sophorolipid, 30g/L of sodium dodecyl benzene sulfonate and 600075g/L of polyethylene glycol;
the R2 reagent includes: anti-human adiponectin antibody latex particles of 9mg/L, pH ═ 7 trihydroxymethyl aminomethane 110mmol/L, tween-8090 g/L and Proclin-3000.3 g/L
Example 4 adiponectin measurement kit
The components and the concentrations thereof are as follows:
the R1 reagent includes: 100mmol/L of trihydroxymethyl aminomethane with pH of 7, 5g/L of sodium chloride, 3g/L of mannitol, 1g/L of chitosan, 12g/L of sophorolipid, 24g/L of sodium dodecyl benzene sulfonate and 600060g/L of polyethylene glycol;
the R2 reagent includes: 10mg/L, pH ═ 7 of latex particles of anti-human adiponectin antibody, 100mmol/L of tris (hydroxymethyl) aminomethane, 80g/L of tween and 3000.2 g/L of Proclin
Comparative example 1
The difference from example 4 is that: the surfactant is sodium dodecyl benzene sulfonate and polyethylene glycol 6000 only, namely the R1 reagent comprises: 100mmol/L of trihydroxymethyl aminomethane with pH value of 7, 5g/L of sodium chloride, 3g/L of mannitol, 1g/L of chitosan, 24g/L of sodium dodecyl benzene sulfonate and 600060g/L of polyethylene glycol; the other steps and operations were the same as in example 4.
Comparative example 2
The difference from example 4 is that: the surfactant is only sophorolipid and polyethylene glycol 6000, i.e. the R1 reagent comprises: 100mmol/L of trihydroxymethyl aminomethane with pH value of 7, 5g/L of sodium chloride, 3g/L of mannitol, 1g/L of chitosan, 12g/L of sophorolipid and 600060g/L of polyethylene glycol; the other steps and operations were the same as in example 4.
Comparative example 3
The difference from example 4 is that: the surfactant is sophorolipid, sodium dodecyl benzene sulfonate and polyethylene glycol 6000 in a mass ratio of 1:1:3, namely the R1 reagent comprises: 100mmol/L of trihydroxymethyl aminomethane with pH value of 7, 5g/L of sodium chloride, 3g/L of mannitol, 1g/L of chitosan, 19.2g/L of sophorolipid, 19.2g/L of sodium dodecyl benzene sulfonate and 600057.6 g/L of polyethylene glycol; the other steps and operations were the same as in example 4.
Comparative example 4
The difference from example 4 is that: the stabilizer is mannitol only, i.e. the R1 reagent includes: the R1 reagent includes: 100mmol/L of trihydroxymethyl aminomethane with pH value of 7, 5g/L of sodium chloride, 4g/L of mannitol, 12g/L of sophorolipid, 24g/L of sodium dodecyl benzene sulfonate and 600060g/L of polyethylene glycol; the other steps and operations were the same as in example 4.
Comparative example 5
The difference from example 4 is that: the concentration ratio of sophorolipid to chitosan is 1:1, i.e. the R1 reagent comprises: the R1 reagent includes: 100mmol/L of trihydroxymethyl aminomethane with pH value of 7, 5g/L of sodium chloride, 3g/L of mannitol, 6.5g/L of chitosan, 6.5g/L of sophorolipid, 24g/L of sodium dodecyl benzene sulfonate and 600060g/L of polyethylene glycol; the other steps and operations were the same as in example 4.
Comparative example 6
The kit was prepared by the method disclosed in prior art CN111239421A, and the anti-human adiponectin antibody latex particles used the purchased components of the present invention.
Effect test
Experimental example 1 Standard Curve of adiponectin measurement kit
The adiponectin standard was used to prepare adiponectin standard samples at concentrations of 0, 10, 12, 14, 16, 18, and 20mg/L, and the adiponectin kit standard curve (shown in FIG. 1) was obtained by the above-described measurement procedure in Experimental example 4 as an example. Wherein, the absorbance change value delta A (A standard-A blank) of the standard substance is a vertical coordinate, the corresponding concentration c of the standard substance is a horizontal coordinate, and a standard curve equation: 0.0099x-0.0002，R2＝0.9998。
Experimental example 2 Linear Range testing
Adiponectin standards were prepared in concentrations of 0.2, 0.5, 1.5, 5.0, 20, 30, and 40mg/L using physiological saline, and the measurement was repeated 10 times per sample of concentration level to obtain an average, and the adiponectin content was measured in mg/L using the kits described in examples 1 to 4 and comparative example 6, respectively, and the results are shown in Table 1 below.
TABLE 1
According to the detection results in the table 2, when the concentration is 0.2-40mg/L, the test results of the kit prepared in the embodiments 1-4 of the present invention can reach the theoretical value and the correlation between the test results and the theoretical value is good, especially the R in the detection of the kit prepared in the embodiment 420.9999 (as shown in fig. 5), the correlation was better than that of the kit (R) prepared in comparative example 620.9992), the kit provided by the invention has a wide linear range.
Experimental example 3 Long-term stability Studies of adiponectin kit
The kits prepared in examples 1 to 4 and comparative examples 1 to 5 prepared by the invention are placed in a dark and sealed condition at 2 to 8 ℃ for 16 months, the detection stability of the kits is detected, 5 times of repeated measurement are carried out on a sample reagent of 5.0mg/L for detection, an average value is obtained, and the average deviation is calculated, which is specifically shown in the following table 2.
TABLE 2 Long term stability test
It can be seen from the test data in table 2 above that the long-term stability of the kits prepared in examples 1-4 of the present invention is significantly higher than that of the kits prepared in comparative examples 1-5, and the comparative examples 4-5 change the type of the stabilizer or change the concentration ratio of sophorolipid and chitosan out of the range disclosed in the present invention, the stability of the kits prepared therefrom is significantly reduced, and the change rate of the test of the kits left for a long time is as high as 30%, which is poor.
EXAMPLE 4 precision test
The adiponectin assay kits prepared in examples 1 to 4 and comparative examples 1 to 5 were repeatedly measured for a plurality of times on the same test specimen at a concentration of 10.0mg/L, and the SD and CV were calculated from the results, and the assay data are shown in Table 3.
TABLE 3
According to the detection data in table 3 above, it can be seen that the precision of the kits prepared in examples 1-4 of the present invention is significantly higher than that of comparative examples 1-5, the coefficient of variation of the kits prepared in example 4 of the present invention is only 1.3%, and is significantly lower than that of other examples, the precision of the kits is significantly reduced when the surfactant component and concentration ratio in reagent R1 are changed in comparative examples 1-3, and the precision of the kits is also affected and reduced when the stabilizer component and concentration ratio are changed in comparative examples 4-5.
Example 5 interference experiments
In normal serum, hemoglobin, triglyceride, bilirubin, and ascorbic acid were each added in a certain amount, and in the same time, deionized water was added in an equal volume as an interferent serum, and the concentration of the sample was measured using the adiponectin measurement kit of examples 1 to 4. The interference degree of 5 percent is taken as the highest tolerance limit of a measuring system to an interfering substance, and the measurement result of adiponectin is not influenced by blood hemoglobin of less than 5.0g/L, triglyceride of less than 10.0g/L, bilirubin of less than 500mg/L and ascorbic acid of less than 10.0g/L in serum.
The reagent used by the invention is simple and easy to obtain, the accuracy, the precision and the stability of the kit are obviously improved by controlling the concentration ratio of various reagents, the cost of the kit is reduced, and the kit is favorable for wide clinical application.
The above-mentioned embodiments are merely exemplary and not intended to limit the present invention, and any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. An adiponectin measurement kit comprising a R1 reagent and a R2 reagent, wherein:
the R1 reagent comprises a buffer solution, an electrolyte, a stabilizer and a surfactant A;
the R2 reagent comprises anti-human adiponectin antibody latex particles, buffer solution, surfactant B and preservative;
the surfactant A is one or more of sophorolipid, sodium dodecyl benzene sulfonate, polyethylene glycol mono-octyl phenyl ether, polyethylene glycol 6000, triton X-100 and polyvinylpyrrolidone.
2. The adiponectin measurement kit according to claim 1, characterized in that: the surfactant is a mixture of sophorolipid, sodium dodecyl benzene sulfonate and polyethylene glycol 6000 in a mass ratio of 1:1-3: 5.
3. The adiponectin measurement kit according to claim 1, characterized in that: the surfactant B is one or more of Tween-20, Tween-80, span-60, span-80 and Triton X-100.
4. The adiponectin measurement kit according to claim 1, characterized in that: the stabilizer is one or more of mannitol, sorbitol, trehalose, sucrose and chitosan.
5. The adiponectin measurement kit according to claim 4, characterized in that: the stabilizer is mannitol and chitosan with the mass ratio of 1-5: 1.
6. The adiponectin measurement kit according to claim 5, characterized in that: the stabilizer is mannitol and chitosan in a mass ratio of 3: 1.
7. The adiponectin measurement kit according to claim 6, characterized in that: the concentration ratio of sophorolipid to chitosan is 5-15: 1.
8. The adiponectin assay kit according to any one of claims 1 to 7, wherein: the adiponectin determination kit is characterized in that the R1 reagent comprises 90-120mmol/L of buffer solution, 3-8g/L of electrolyte, 2-5g/L of mannitol, 1-3g/L of chitosan, 10-15g/L of sophorolipid, 20-40g/L of sodium dodecyl benzene sulfonate and 600050-80 g/L of polyethylene glycol;
the R2 reagent comprises 8-12mg/L of anti-human adiponectin antibody latex particles, 90-120mmol/L of buffer solution, 100g/L of Tween-8060 and 0.1-0.5g/L of preservative.
9. The adiponectin assay kit according to any one of claims 1 to 7, wherein: the adiponectin determination kit is characterized in that the R1 reagent comprises 90-120mmol/L of trihydroxymethyl aminomethane with pH of 7, 3-8g/L of sodium chloride, 2-5g/L of mannitol, 1-3g/L of chitosan, 10-15g/L of sophorolipid, 20-40g/L of sodium dodecyl benzene sulfonate and 600050-80 g/L of polyethylene glycol;
the R2 reagent comprises 90-120mmol/L of trihydroxymethyl aminomethane of anti-human adiponectin antibody latex particles of 8-12mg/L, pH ═ 7, 100g/L of Tween-8060-.
10. The adiponectin assay kit according to any one of claims 1 to 7, wherein: the adiponectin determination kit is characterized in that the R1 reagent comprises 100mmol/L of trihydroxymethyl aminomethane with pH of 7, 5g/L of sodium chloride, 3g/L of mannitol, 1g/L of chitosan, 12g/L of sophorolipid, 24g/L of sodium dodecyl benzene sulfonate and 600060g/L of polyethylene glycol;
the R2 reagent comprises anti-human adiponectin antibody latex particles of 10mg/L, pH ═ 7 of tris (hydroxymethyl) aminomethane 100mmol/L, Tween-8080 g/L and Proclin-3000.2 g/L.
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| CN114544527A (en) * | 2022-01-26 | 2022-05-27 | 浙江夸克生物科技有限公司 | Saliva liquefaction sugar chain antigen KL-6 determination kit and preparation method thereof |
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