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Test kit for measuring content of retinol conjugated protein in serum of human body and preparation method of test kit

A technology that binds proteins and retinol, and is applied in measurement devices, biological tests, material inspection products, etc., to achieve the effects of good stability, improved efficiency and good specificity

Inactive Publication Date: 2017-07-07
SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, clinical testing requires the kit to meet the requirements of strong specificity, high sensitivity, strong anti-interference, wide linear range, and good bottle opening stability. Therefore, these technical issues need further in-depth research.

Method used

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  • Test kit for measuring content of retinol conjugated protein in serum of human body and preparation method of test kit
  • Test kit for measuring content of retinol conjugated protein in serum of human body and preparation method of test kit
  • Test kit for measuring content of retinol conjugated protein in serum of human body and preparation method of test kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Preparation of kit for measuring human serum retinol-binding protein content in the present invention

[0043] Preparation of reagent R1: get 995g water, then add 2.4g sodium dihydrogen phosphate, 11.4g disodium hydrogen phosphate, 10.0g polyethylene glycol 20000, 9.0g NaCl, 0.5g Tween 20, 5.0g bovine serum albumin ( w / w) and 0.2g Proclin300, each addition of material needs to be stirred until it is completely dissolved, and the pH is adjusted to 7.4; filter with a 0.22μm filter membrane to prepare 1L of reagent R1.

[0044] Preparation of reagent R2: 0.3% polystyrene latex particles (w / w) bound to retinol-binding protein antibody, 1% NaCl (w / w), 0.05% Tween 20 in 50mmol / L Mops buffer (w / w), 1% BSA (w / w), 0.02% Proclin 300 (w / w). Among them, the antibodies coupled to the latex microspheres of two particle sizes are polyclonal antibodies of rabbit anti-human retinol binding protein, and the antibodies with different particle sizes are prepared separately and mixed at a ...

Embodiment 2

[0057] Preparation of kit for measuring human serum retinol-binding protein content in the present invention

[0058] Preparation of reagent R1: get 995g water, then add 2.4g sodium dihydrogen phosphate, 11.4g disodium hydrogen phosphate, 15.0g polyethylene glycol 20000, 9.0g NaCl, 0.8g Tween 20, 5.0g bovine serum albumin ( w / w) and 0.2g Proclin300, each addition of material needs to be stirred until it is completely dissolved, and the pH is adjusted to 7.2; filter with a 0.22μm filter membrane to prepare 1L of reagent R1.

[0059] Preparation of reagent R2: 0.3% polystyrene latex particles (w / w) bound to retinol-binding protein antibody, 1% NaCl (w / w), 0.05% Tween 20 in 50mmol / L Mops buffer (w / w), 1% BSA (w / w), 0.02% Proclin 300 (w / w). Among them, the latex particles with small particle size are coupled with mouse anti-human retinol binding protein monoclonal antibody, and the latex particles with large particle size are coupled with rabbit anti-human retinol binding protein...

Embodiment 3

[0072] Performance test of the kit (made in Example 1) for measuring retinol-binding protein of the present invention

[0073] Analytical method of performance test: two-point endpoint method. Take 225ul of reagent R1, add 3ul of sample, add 75ul of reagent R2 after 5min at 37°C, start reading, react for 5min, read the point again, and get the absorbance difference (△A).

[0074] Calculation method: multi-point calibration, a calibration curve is made according to the absorbance and reference serum values, and the standard curve is used as the calculation mode, and the sample content can be calculated on the curve according to its absorbance.

[0075] Reference range: 25-70mg / L

[0076] Testing instrument: Hitachi 7100 automatic biochemical analyzer.

[0077] Detection wavelength: 600nm.

[0078] 1. Linearity Test of Standard Serum Samples

[0079] Dissolve the natural retinol-binding protein in a solution similar to human serum matrix (0.9% NaCl, 0.1% bovine serum albumin...

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Abstract

The invention provides a test kit for measuring the content of retinol conjugated protein in serum of a human body and a preparation method of the test kit. The volume ratio of a reagent R1 and a reagent R2 is 3:1. The R1 comprises 0.01-4%w / w of emulsifiers, 0.1-2%w / w of sodium chloride, 0.05-2%w / w of sensitizers, 0.02%w / w of preservative and 30-100mmol / L of buffer solutions. The R2 comprises 0.1-0.5%w / w of latex particle combined with a retinol conjugated protein antibody, 0.1-1.0%w / w of bovine serum albumin, 0.02-2%w / w of Tween 20, 0.02-2%w / w of stabilizers 2, 0.02-2%w / w of protein protective agents, 0.02%w / w of preservative and 20-50mmol / L of buffer solutions. The test kit has wide linearity range, high sensitivity and stability and good clinical application prospect.

Description

technical field [0001] The invention relates to a biological reagent, in particular to a detection kit, in particular to a stable latex immunoturbidimetric method for measuring the content of retinol-binding protein in human serum and a preparation method thereof. Background technique [0002] Retinol binding protein (RBP) is a protein with a single peptide chain, containing 183 amino acid residues, with a relative molecular weight of about 21,000 and a half-life of about 3 to 12 hours. It belongs to the Lipocalin protein superfamily member. RBP is a transporter of retinol (vitamin A) in the blood. It is synthesized by the liver and widely distributed in blood, cerebrospinal fluid, urine and other body fluids. Its main function is to transport retinol (vitamin A) from liver cells to the epithelium. cell. It is now believed that RBP in the blood mainly exists in the form of a complex bound to retinol and prealbumin. When retinol in the complex binds to target cells, RBP faci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/544
CPCG01N33/68G01N33/544
Inventor 张龙平景盛
Owner SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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