Wider linear range retinol binding protein determination kit

A protein-binding and linear range technology, applied in the field of medical testing, can solve the problems of short reaction time, detection sensitivity and linear range to be further improved, and good precision, achieving high accuracy, improved detection sensitivity, and good precision Effect

Inactive Publication Date: 2015-05-13
JIANGSU HAOSHEN MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PETIA detection method has short reaction time, good precision, wide detection range, and is not affected by jaundice, hemolysis and lipemia specimens. It is ea...

Method used

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  • Wider linear range retinol binding protein determination kit
  • Wider linear range retinol binding protein determination kit
  • Wider linear range retinol binding protein determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of Retinol Binding Protein Assay Kit

[0023] 1. Preparation of Reagent R1

[0024] Add final concentrations of 2% polyethylene glycol 6000, 0.8% NaCl, 0.1% bovine serum albumin, and 0.1% sodium azide to 20 mmol / L Tris-HCl buffer (pH 7.4), and mix well. Evenly, filter with a 0.22 μm filter membrane to make reagent R1.

[0025] 2. Preparation of Reagent R2

[0026] Mouse anti-human retinol binding protein monoclonal antibody and rabbit anti human retinol binding protein polyclonal antibody are both entrusted to professional companies to produce according to conventional methods. The antibody only reacts with human retinol binding protein and has no immunity to other antigens Cross-reactivity and potency can meet the needs of this test.

[0027] Add EDAC:Sulfo-NHS:nanolatex particles (250nm or 60nm) to 20mmol (pH6.1) NES buffer solution, the mass ratio is 1:0.6:1, mix well, activate on a rotary shaker at room temperature for 1 hour, and centrifuge at 17,0...

Embodiment 2

[0030] Performance Test of Retinol Binding Protein Assay Kit

[0031] Experimental analysis method: two-point endpoint method. Take 240 ul of reagent R1, add 3 ul of sample, add 60 ul of reagent R2 after 5 minutes at 37°C, start to read the point, react for 5 minutes, read the point again, and obtain the absorbance difference (△A).

[0032] Testing instrument: Hitachi 7180 automatic analyzer.

[0033] Detection wavelength: the main wavelength is 600nm, and the secondary wavelength is 800nm.

[0034] 1. Sensitivity test

[0035] Detect water 20 times, record the absorbance difference (△A 水 ), calculate the mean value (X 水 ) and standard deviation (SD 水 ); detect the sample with a concentration of 30mg / L 20 times, and record the absorbance difference (△A 样 ), calculate the mean value (X 样 ) and standard deviation (SD 样 ), and the test results are shown in Table 1. Calculate the sensitivity using the following formula.

[0036] Sensitivity = 30 × (X 水 +3×SD 水 ) / X ...

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Abstract

The invention provides a wider linear range retinol binding protein determination kit including reagent R1 and reagent R2. The reagent R1 is a reaction liquid for promoting the antibody antigen specific combination; the reagent R2 is a buffer of nano latex particles combined with anti human retinol binding protein antibody. The reagent R2 is characterized in that: polystyrene nano latex particles with the diameter of 50-300 nm are coupled with retinol binding protein monoclonal antibody or polyclonal antibody to obtain conjugates with different particle sizes and different antibodies, the conjugates with different particle sizes and different antibodies are mixed in different proportions to prepare the reagent R2 for detection of human serum retinol binding protein, and the detection sensitivity and the linear range of the kit can be improved. The detection sensitivity of the kit is 0.01mg/ L, and the linear range is 0-150 mg/ L.

Description

technical field [0001] The invention belongs to the field of medical testing, and in particular relates to a nano-latex microparticle type retinol-binding protein assay kit. Background technique [0002] Retinol binding protein (RBP) is a single peptide chain protein, containing 183 amino acid residues and 3 disulfide bonds, with a relative molecular weight of 21,000 and a half-life of about 3-12 hours. family members. 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 from liver cells to epithelial cells. RBP in the blood mainly exists in the form of complexes bound to retinol and prealbumin. When retinol in the complex binds to target cells, RBP is separated from prealbumin, filtered out from the glomerulus, and released by the proximal kidney. Absorption and degradation by tubular epithelial cells. [0003] I...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/68
CPCG01N33/577G01N33/68
Inventor 董同义李玲董海江张文静
Owner JIANGSU HAOSHEN MEDICAL TECH
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