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Kit for performing retinol binding protein detection by using latex turbidimetry

A technology that combines protein and latex turbidity, and is used in measurement devices, color/spectral property measurements, instruments, etc.

Inactive Publication Date: 2013-02-27
BEIJING KANGMEI TIANHONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to overcome the deficiencies of the existing RBP detection kits at present, and provide a latex-enhanced immune turbidimetric kit for detecting RBP with high sensitivity

Method used

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  • Kit for performing retinol binding protein detection by using latex turbidimetry
  • Kit for performing retinol binding protein detection by using latex turbidimetry
  • Kit for performing retinol binding protein detection by using latex turbidimetry

Examples

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Embodiment 1

[0026] In order to make the present invention easier to understand, the present invention will be further described below in conjunction with specific examples, and these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Example 1: Preparation of Retinol Binding Protein Latex Immunization Kit

[0027] 1. Preparation of reagent R1

[0028] Dissolve NaCl (7.0-9.0g) in double-distilled water first, then add Tris-HCl buffer solution, and finally add double-distilled water to 1000ml, so that the final concentration of Tris is 0.05mol / L, shake well, then add a small amount of PEG 6000 and Tween80, just mix well.

[0029] Those skilled in the art can also choose other conventional buffers, such as one or more of phosphate buffer, HEPES buffer, glycine buffer, and barbiturate buffer.

[0030] 2. Preparation of reagent R2

[0031] Antibody mixture preparation: Take 2ml of 1mg / ml anti-retinol-binding protein m...

Embodiment 2

[0035] Embodiment 2: Assay method of retinol binding protein detection reagent

[0036] This kit is suitable for fully automatic or semi-automatic biochemical analyzers of Beckman, Hitachi, Olympus, Toshiba, Roche, Abbott, Siemens, Mindray and other brands. The determination method is as follows:

[0037] 1. Measurement conditions

[0038] The assay condition of kit of the present invention

[0039] Temperature: 37°C

[0040] Dominant wavelength: 560nm

[0041] Secondary wavelength: 800nm

[0042] Sample volume: 5 μL

[0043] R1 dosage: 200μL

[0044] R2 dosage: 40μL

[0045] Analysis type: Two-point endpoint method

[0046] 2. The measurement method is as follows:

[0047] To measure the absorbance of the blank, add 5 μL sample to 200ul R1 reagent, pre-incubate for 5 minutes, measure the absorbance of the blank, add 40 μL R2 reagent, incubate for 5 minutes, and measure the absorbance of the reaction.

[0048] The sample volume, R1 dosage and R2 dosage can be adjusted...

Embodiment 3

[0051] Example 3: Analytical Performance Evaluation of Retinol Binding Protein Immunological Reagents

[0052] 1. Analytical sensitivity or minimum detection limit

[0053] Control reagent: a well-known foreign brand

[0054] Detection principle: Use latex-enhanced immunoturbidimetry to detect the change of turbidity at 570nm, and the change degree is linearly related to the RBP content in the sample.

[0055] Sensitivity evaluation method: Dilute the calibrator into a series of concentrations close to the sensitivity, and determine its concentration by ELISA method to be 0.05mg / L, 0.10mg / L, 0.15mg / L, 0.21mg / L, add zero standard as the sample, Using two kits, according to their respective measurement methods, repeat the measurement 10 times, and record the reaction absorbance value:

[0056] Table 1: Sensitivity measured value of embodiment 1

[0057]

[0058] The average value of the zero standard for 10 repeated determinations is -0.1, the standard deviation SD is 2.81...

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Abstract

The invention relates to the technical field of biotechnology, and particularly discloses a kit for detecting retinol binding protein content by using latex immunoturbidimetry. The kit comprises a reagent R1, a reagent R2 and a standard product, wherein the reagent R1 is buffer solution with pH (Potential of Hydrogen) value of 6-9; the reagent R2 is latex reagent coated by anti-retinol binding protein double antibodies; and the standard product is retinol binding protein solution with pH value of 5-8. According to the kit, the retinol binding protein content in a sample can be detected by using the latex immunoturbidimetry; the sensitivity is high and can reach 0.042 mg / L; and the kit has the advantages of high stability, easiness and quickness in operation, high specificity, low probability of interference, accurate quantification and broad application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting retinol-binding protein by using double antibodies to prepare latex immune reagents and using latex immune turbidimetry. Background technique [0002] Retinol binding protein (RBP) is a single peptide chain protein with a molecular weight of about 21kD. It has a site that specifically binds a molecule of all-trans retinol, namely vitamin A. Retinol-binding proteins can be divided into five categories according to sequence homology: secreted RBP, intracellular RBP, nuclear receptor for retinoic acid, visual tissue-specific extracellular RBP and visual tissue-specific intracellular RBP. Serum RBP4 is a secreted RBP, which is the main extracellular transporter responsible for binding and transporting retinol in the blood. [0003] Retinol-binding protein is a vitamin transporter in the blood, synthesized by the liver, and widely distributed in human serum, cerebrosp...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/31
Inventor 张英伟
Owner BEIJING KANGMEI TIANHONG BIOTECH
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