Reagent for determining content of human cholyglycine by using latex immunoturbidimetry technology

A technology of latex immunity and glycocholic acid, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor stability of labeled enzymes and cannot meet clinical use, and achieve the advantages of reagent stability and increase sensitivity

Inactive Publication Date: 2018-12-11
北京安图生物工程有限公司
View PDF11 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the poor stability of the current labeling enzymes, they cannot meet the requirements of clinical use.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reagent for determining content of human cholyglycine by using latex immunoturbidimetry technology
  • Reagent for determining content of human cholyglycine by using latex immunoturbidimetry technology
  • Reagent for determining content of human cholyglycine by using latex immunoturbidimetry technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Prepare the reagent of mensuration human serum glycocholic acid content by the method of the present invention

[0026] 1. Preparation of reagents 1

[0027] 1. Preparation of latex microspheres-BSA-glycocholic acid conjugates

[0028] 1) Activation of Glycocholic Acid

[0029] Accurately weigh 400mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), dissolve it in 100mL MES buffer (20mmol / L, pH=6.0) to obtain EDC solution; Accurately weigh 200mg glycocholic acid and 400mg Sulfo-NHS, dissolve in 100mL MES buffer (20mmol / L, pH=6.0) to obtain a glycocholic acid solution; slowly add the EDC solution to the glycocholic acid solution while stirring After the addition, the stirring reaction was continued for 30 minutes to obtain an activated glycocholic acid solution.

[0030] 2) Coupling of Glycocholic Acid and BSA

[0031] Accurately weigh 100mg of BSA and dissolve it in 100mL MES buffer (20mmol / L, pH=6.0) to obtain the BSA solution; while stirring, sl...

Embodiment 2

[0040] Embodiment 2 detection method and equipment

[0041] The detection of glycocholic acid in the present invention requires fresh serum or plasma without hemolysis. Samples that interfere with the reaction absorbance, such as hemolysis or lipid turbidity, may affect the final result. It is recommended to re-collect blood.

[0042] Reagent of the present invention is applied to various types of biochemical instruments, such as Hitachi Hitachi7080 / 7600P, Olympus Olympus AU400, Beckman Coulter LX20 / AU680 / AU5800, Toshiba Toshiba 120FR, Abbott C8000, Siemens SIEMENS ADVIA1800 / ADVIA2400 / Dimension RxL Max biochemical analyzer, etc.

[0043] The specific detection parameter settings are shown in the table below, and the reading time of A1 and A2 is set according to different instruments:

[0044]

Embodiment 3

[0045] Embodiment 3 correlation comparison

[0046] With the reagent prepared in Example 1, the same sample is detected with the routinely used radioimmunoassay and homogeneous enzyme immunoassay, and the correlation data obtained are shown in the following table 1 and figure 1 :

[0047]

[0048] We can find out from Table 1 that the measured data of the present invention is close to the result obtained by radioimmunoassay, and the measured result of homogeneous enzyme immunoassay is better correlated with the result obtained by radioimmunoassay in the low value stage, but the high value stage is significantly On the low side, so its linearity is lower than that of the present invention and the radioimmunoassay reagent.

[0049] figure 1 Then relatively intuitively embodies the comparative relationship of the three reagents, the trend line of the measured data of the present invention and the result obtained by radioimmunoassay is close to a straight line, showing that b...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to view more

Abstract

The invention discloses a reagent for determining the content of human cholyglycine by using a latex immunoturbidimetry technology. The reagent is prepared from a reagent body 1 and a reagent body 2,wherein the reagent body 1 is prepared by adding 100 mM of an auxiliary agent, a phosphate buffer solution with the pH value of 8.0, a sodium chloride solution with the concentration of 0.9%, BSA withthe concentration of 0.1% and a stabilizer into a latex microsphere-BSA-cholyglycine conjugate with the concentration of 0.04%, and the reagent body 2 is prepared by adding 15 mM of an auxiliary agent, a phosphate buffer solution with the pH value of 7.4, a sodium chloride solution with the concentration of 0.9%, BSA with the concentration of 0.1%, a surfactant with the concentration of 0.1% anda stabilizer into an anti-mouse cholyglycine monoclonal antibody with the concentration of 5%. According to the reagent, latex microspheres are introduced into the reagent, due to the existence of thelatex microspheres, the sensitivity of the detection reagent is greatly improved, and the requirements of clinical use are met. Compared with homogeneous enzyme immunoassay, the reagent has great advantages in stability, under the acceleration condition, and the stability time of the reagent is at least 2 times or above of the stability time of a homogeneous enzyme immunoassay reagent.

Description

technical field [0001] The invention relates to biological detection reagents, in particular to a reagent for measuring glycocholic acid content in human serum by using latex immunoturbidimetric technology. Background technique [0002] Serum glycocholic acid (Cholyglycine, CG, molecular weight 465.6) is a kind of bile acid. Bile acids include a large class of cholic acids. In bile, there are mainly cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), and a small amount of lithocholic acid (LCA) and trace amounts of ursodeoxycholic acid. Cholic acid (UDCA), all kinds of cholic acid may be combined with glycine and taurine to form a combined form, which is the main form secreted by the liver into bile. All kinds of bile acids, whether they are free or bound, contain both hydrophilic groups (hydroxyl, carboxyl, sulfonyl) and hydrophobic groups (methyl and hydrocarbon core) inside their molecules, so the three-dimensional structure of bile acids The configu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 刘泊湾
Owner 北京安图生物工程有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap