Retinol binding protein determination kit and preparation method thereof

A protein and retinol-binding technology, applied in the fields of medicine and biochemistry, can solve the problems of complex operation, radioactive contamination, and poor accuracy, and achieve the effects of increased solubility, high accuracy, and rapid response

Inactive Publication Date: 2016-11-09
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a kit for measuring retinol binding protein and its preparation method in order to overcome the defects of complex operation, radioactive contamination and poor accuracy in the prior art

Method used

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  • Retinol binding protein determination kit and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0074] Reagent R1:

[0075] Tris buffer 100 mmol / L

[0076] Dithizone 30 mmol / L

[0077] Polyethylene glycol-8000 20 g / L

[0078] Bovine Serum Albumin 40 g / L

[0079] Sodium azide 0.7 g / L

[0080] The solvent is purified water

[0081] Reagent R2:

[0082] Tris buffer 100 mmol / L

[0083] Bovine serum albumin 12g / L

[0084] Triton-100 0.7 mL / L

[0085] Glycerin 45 mmol / L

[0086] Sodium azide 0.7g / L

[0087] Latex coated anti-human retinol binding protein antibody 4g / L

[0088] Its solvent is purified water.

Embodiment 2

[0090] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0091] Reagent R1:

[0092] PBS buffer 150 mmol / L

[0093] 8-hydroxyquinoline 10 mmol / L

[0094] Polyethylene glycol-6000 30 g / L

[0095] Sucrose 20g / L

[0096] Phenol 1.1 g / L

[0097] The solvent is purified water

[0098] Reagent R2:

[0099] PBS buffer 50mmol / L

[0100] Bovine serum albumin 18 g / L

[0101] Triton-100 1.0 mL / L

[0102] Sodium alginate 60 mmol / L

[0103] Phenol 0.3 g / L

[0104] Latex coated anti-human retinol binding protein antibody 8g / L

[0105] Its solvent is purified water.

Embodiment 3

[0107] Kit preparation and method of use

[0108] 1. Preparation of latex-coated anti-human retinol-binding protein antibody: Dilute polystyrene microspheres with a particle size of 120 nm with 50 mmol / L MES buffer solution to obtain a mass concentration of polystyrene microspheres of 3% solution, and then add 0.5 mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride to each milliliter of the solution, react at 25°C for 2 hours, use Centrifuge at 15,000 rpm / min for 30 minutes, remove the supernatant, suspend the pellet in 50 mmol / L MES buffer, ultrasonically disperse, and then use a centrifuge to centrifuge at 15,000 rpm / min for 30 minutes Minutes, remove the supernatant, suspend the precipitate in 50 mmol / L MES buffer, make the final mass concentration of polystyrene microspheres 2%, ultrasonically disperse, and then add an equal volume of anti-human retinol containing The MES buffer solution bound to the protein antibody was mixed and stirred, and reacted at 25...

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Abstract

The invention discloses a retinol binding protein determination kit and a preparation method thereof. The kit comprises a reagent R1 and a reagent R2 which are liquid components and are mutually independent, the reagent R1 comprises a buffer solution, a chelating agent, an accelerator, a stabilizer and an antiseptic, and the reagent R2 comprises the buffer solution, a stabilizer, a surfactant, a suspending aid, the antiseptic and a latex coated anti-retinol binding protein antibody. The preparation method comprises the following steps: preparing the reagents according to the component content; mixing a sample to be determined with the reagent R1 and the reagent R2 to fully react the sample to be determined with the reagent R1 and the reagent R2; determining the absorbance difference by using a fully-automatic biochemical analyzer after the reaction; and calculating the concentration of the retinol binding protein in the sample according to the absorbance change value. The kit has high accuracy.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring retinol-binding protein and a preparation method thereof. Background technique [0002] The Retinol-Binding Protein (RBP) molecule is composed of a polypeptide chain consisting of 183 amino acid residues and a small part of sugar, without neutral sugar and hexosamine, with a molecular weight of 21000 and a sedimentation coefficient of 3S , the isoelectric point is pH 4.4~4.8, and the half-life is 3~12 hours. The retinol-binding protein was subjected to ethyl dextran chromatography to obtain two peaks, A and B, and diethylaminoethanol dextran chromatography to obtain A1, A2 and B1, B2 four parts. The immunodiffusion method showed that the antigenicity of the four parts of retinol binding protein was consistent. Retinol binding protein has three forms, namely RBP, RBP1, RPB2, the leucine residue at the C-terminal of the polypeptide chain of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/544G01N33/543G01N33/536G01N33/531G01N21/75
CPCG01N33/6893G01N21/75G01N33/531G01N33/536G01N33/54313G01N33/544
Inventor 蔡晓辉庄庆华吴铮徐运
Owner ANHUI IPROCOM BIOTECH CO LTD
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