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Free protein S determination kit and preparation method thereof

A kit and reagent technology, applied in the field of immunoassay, can solve the problems of poor coagulation method specificity, high measurement value, false positive, etc., and achieve the effect of improving sensitivity and specificity, improving sensitivity and high accuracy

Inactive Publication Date: 2022-08-02
SHENZHEN DYMIND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Most of the free protein S (FPS) assay kits currently available on the market are protein S activity detection kits using the coagulation method. In the case of excessive content, false positives will occur, resulting in abnormally high measured values

Method used

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  • Free protein S determination kit and preparation method thereof
  • Free protein S determination kit and preparation method thereof
  • Free protein S determination kit and preparation method thereof

Examples

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preparation example Construction

[0051] A preparation method of the above-mentioned test kit, comprising the following steps:

[0052] S1. Add the first stabilizer and the coagulant to the first buffer, adjust the pH to 6.0-9.0, and prepare the R1 reagent.

[0053] S2. The anti-human FPS antibody latex particles and the second stabilizer are added to the second buffer, and the pH is adjusted to 6.0-9.0 to prepare the R2 reagent.

[0054] Further, preferably in S2, it specifically includes the following steps: S21, taking latex particles and adding them to the activation buffer, adding an activator for activation, to obtain activated latex particles; specifically, the activation buffer is an MES solution, and the activation of the latex particles The agent is EDC, the ratio of latex particles to activator is 1:(2~20), and the activation time is 20~40min. Anti-human FPS antibody is added to the activated latex particles for coupling, and the reaction time is 2~4h. Anti-human FPS antibody latex particles, after...

Embodiment 1

[0057] A free protein S assay kit, including R1 reagent and R2 reagent; the specific components are shown in the following table:

[0058] Table 1. Reagent components of the kit of Example 1

[0059]

[0060] Note: " / " means that this ingredient is not included.

[0061] The preparation method of the kit includes the following steps:

[0062] R1 reagent: add 1% sodium chloride, 2% dextran 8000 and 0.03% PC300 to 100 mL of imidazole buffer, adjust the pH to 7.0, and mix well to obtain R1 reagent.

[0063] R2 reagent: use an ultrafiltration tube to wash the anti-human FPS antibody; use MES solution with pH value of 6.5 to dilute the carboxypolystyrene latex particles with a particle size of 180 nm to a final concentration of 0.5%; add 10% activator EDC (1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride)) solution into the above solution, the ratio of latex particles and activator is 1:2, incubate for 30 minutes, add affinity Chromatographic anti-human FPS antibody...

Embodiment 2

[0065] A free protein S assay kit, including R1 reagent and R2 reagent; the specific components are shown in the following table:

[0066] Table 2. Recipe of each component of the kit of Example 2

[0067]

[0068] Note: " / " means that this ingredient is not included.

[0069] The preparation method of the kit includes the following steps:

[0070] R1 reagent: Add 0.5% sodium chloride, 1% dextran 6000 and 0.03% PC300 to 100 mL of imidazole buffer, adjust the pH to 7.0, and mix well to obtain R1 reagent.

[0071] R2 reagent: use an ultrafiltration tube to wash the anti-human FPS antibody; use MES solution with pH value of 6.5 to dilute the carboxypolystyrene latex particles with a particle size of 180 nm to a final concentration of 0.5%; add 10% activator EDC (1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride)) solution into the above solution, the ratio of latex particles and activator is 1:2, incubate for 30 minutes, add affinity Chromatographic anti-human FP...

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Abstract

The invention discloses a free protein S determination kit and a preparation method thereof. The kit comprises a reagent R1 and a reagent R2, the reagent R1 comprises a first buffer solution, a first stabilizer and a coagulant, and the pH value of the reagent R1 is 6.0-9.0; the R2 reagent comprises anti-human FPS antibody latex particles, a second buffer solution and a second stabilizer, the pH value of the R2 reagent is 6.0-9.0, and FPS is free protein S; the preparation method comprises the following steps: adding a first stabilizer and a coagulant into a first buffer solution, and adjusting the pH value to 6.0-9.0 to prepare a reagent R1; adding anti-human FPS antibody latex particles and a second stabilizer into a second buffer solution, and adjusting the pH value to 6.0-9.0 to prepare a reagent R2; according to the invention, the anti-human FPS antibody is coupled with the latex particles, and the FPS in the human plasma is determined by using the latex enhanced immunoturbidimetry, so that the accuracy is high, the stability is good, and the linear range is wide; the preparation method is simple and stable in process.

Description

technical field [0001] The invention relates to the technical field of immunoassays, in particular to a free protein S determination kit and a preparation method thereof. Background technique [0002] Plasma protein S (PS) is a vitamin K-dependent non-enzymatic plasma glycoprotein that exists in plasma in two forms: 40% in free (FPS) form with APC cofactor activity; 60% in plasma PS forms a complex with complement C4b-binding protein (C4bp) in the form of non-covalent bond, and the auxiliary activity of PS in the bound form is very low. There is a dynamic balance between free and bound forms in plasma and can be converted into each other, and the two are collectively referred to as total PS (tPS). Protein S is a physiological anticoagulant substance in the body. During the coagulation process, activated coagulation factors VIII (FVIIIa) and V (FVa) are the rate-limiting factors activated by coagulation factor X (FX) and prothrombin, while the protein C system It can inacti...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/546G01N33/577
CPCG01N33/6893G01N33/54346G01N33/54393G01N33/577G01N2333/47G01N2800/224
Inventor 胡彦勇
Owner SHENZHEN DYMIND BIOTECH
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