Sample diluent for ELISA detection kit and preparation method of sample diluent

A technology of sample diluent and detection kit, which is applied in the field of sample diluent and its preparation of ELISA detection kit, which can solve the problems of inability to effectively protect the sample to be tested and affect the accuracy and repeatability of the test results, and achieve the elimination of Effects of non-specific adsorption, improved detection sensitivity, improved accuracy and repeatability

Pending Publication Date: 2021-06-25
武汉菲恩生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the sample diluent used in the industry is generally PBST solution, that is, PBS-T solution with 0.5% BSA. The advantage of this solution is that the production cost is low, and the disadvantage is that it only has salt buffer solution and cannot effectively protect the sample to be tested. And it is quite different from the serum environment, and the matrix effect is obvious. The application effect of the sample types with low protein concentration such as cell culture supernatant, tissue grinding fluid, and tissue lysate is still acceptable. The recovery rate of standard addition is usually 80%- 120%
However, for samples with high protein and complex components such as serum and plasma, the recovery rate of the spiked standard is only 20%-50%, which seriously affects the accuracy and repeatability of the test results.

Method used

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  • Sample diluent for ELISA detection kit and preparation method of sample diluent
  • Sample diluent for ELISA detection kit and preparation method of sample diluent
  • Sample diluent for ELISA detection kit and preparation method of sample diluent

Examples

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Effect test

Embodiment 1

[0029] A sample diluent for the ELISA detection kit, calculated according to mass percentage, including 2% bovine serum albumin, 0.1% animal IgG, 5% maltose, 0.3% imidazole, 0.5% salt buffer, 0.05% Tween 20 and 0.03% Proclin300, the balance is purified water.

[0030] Animal IgG was obtained after purification of the negative serum ProteinAG of healthy mice; the salt buffer contained 12 g of anhydrous sodium dihydrogen phosphate and 9 g of sodium chloride per liter of salt buffer, and the solvent was purified water.

[0031] A preparation method for a sample diluent of an ELISA detection kit, comprising the steps of:

[0032] S1, weigh 12g of anhydrous sodium dihydrogen phosphate and 9g of sodium chloride into 500ml of purified water, stir until completely dissolved, and obtain salt buffer A;

[0033] S2, weigh 5g imidazole and 40g maltose, add in the salt buffer A in turn, stir for 3min, obtain solution B after dissolving;

[0034] S3, weigh 8g of bovine serum albumin and a...

Embodiment 2

[0037] A sample diluent for ELISA detection kit, calculated according to mass percentage, including 5% bovine serum albumin, 0.3% animal IgG, 8% arginine, 1.5% citric acid, 0.5% salt buffer, 0.1% Tween 20 and 0.18% Proclin300, the balance is purified water.

[0038] Animal IgG is obtained after purification of healthy rabbit negative serum ProteinAG; the salt buffer contains 12 g of anhydrous sodium dihydrogen phosphate and 9 g of sodium chloride per liter of salt buffer, and the solvent is purified water.

[0039] A preparation method for a sample diluent of an ELISA detection kit, comprising the steps of:

[0040] S1, weigh 12g of anhydrous sodium dihydrogen phosphate and 9g of sodium chloride into 700ml of purified water, stir until completely dissolved, and obtain salt buffer solution A;

[0041] S2, weigh 8g of citric acid and 60g of arginine, add them to the salt buffer A in turn, stir for 5min, and obtain solution B after dissolving;

[0042] S3, weigh 13g of rabbit s...

Embodiment 3

[0045] A sample diluent for ELISA detection kit, calculated according to mass percentage, including 4% bovine serum albumin, 0.2% animal IgG, 6% arginine, 1.0% imidazole, 0.5% salt buffer, 0.08% Tween 20 and 0.10% Proclin300, the balance is purified water.

[0046] Animal IgG was obtained after purification of healthy sheep negative serum ProteinAG; the salt buffer contained 12 g of anhydrous sodium dihydrogen phosphate and 9 g of sodium chloride per liter of salt buffer, and the solvent was purified water.

[0047] A preparation method for the sample diluent of ELISA detection kit, is characterized in that, comprises the steps:

[0048] S1, weigh 12g of anhydrous sodium dihydrogen phosphate and 9g of sodium chloride into 600ml of purified water, stir until completely dissolved, and obtain salt buffer A;

[0049]S2, weigh 6g of metal ion exchanger and 50g of protein stabilizer, add them into salt buffer A in sequence, stir for 4min, and obtain solution B after dissolving;

...

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Abstract

The invention provides a sample diluent for an ELISA detection kit. The sample diluent comprises serum albumin, animal IgG, a protein stabilizer, a metal ion exchanger, a salt buffer solution, Tween 20, Proclin300 and purified water. According to the invention, the protein stabilizer and the metal ion exchanger are added into the sample diluent, so that interference of a matrix effect can be effectively reduced, and non-specific adsorption can be removed; and healthy animal IgG is added, so that a to-be-detected sample can be protected, a serum environment can be simulated, interference of heterotropism antibodies and rheumatoid factors in a serum and plasma sample can be reduced, false positive is reduced, and the accuracy and repeatability of ELISA detection results are improved.

Description

technical field [0001] The invention relates to the field of enzyme-linked immunoassay, in particular to a sample diluent used in an ELISA detection kit and a preparation method thereof. Background technique [0002] ELISA (Enzyme-Linked Immunosorbent Assay) is a method for quantifying soluble target proteins in a sample. The sample diluent is one of the core components of the ELISA kit, and its function is to ensure the accuracy of the ELISA kit. Theoretically, the composition of the sample diluent should be exactly the same as that of the sample to be tested except that it does not contain the target protein, but in reality this is impossible, especially for human serum and plasma samples. [0003] At present, the sample diluent used in the industry is generally PBST solution, that is, PBS-T solution with 0.5% BSA. The advantage of this solution is that the production cost is low, and the disadvantage is that it only has salt buffer solution and cannot effectively protect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N1/38
CPCG01N1/38G01N33/543
Inventor 董垚张丽媛
Owner 武汉菲恩生物科技有限公司
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