Preparation method for immunomagnetic microsphere, Sudan red detection kit containing immunomagnetic microsphere and detection method

A technology of magnetic microspheres and Sudan red, which is applied in the field of analysis and detection, can solve the problems of poor stability and low coupling rate, and achieve the effects of good stability, increased coupling rate and short detection time

Active Publication Date: 2015-05-20
BEIJING PURKINJE GENERAL INSTR
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the defects of low coupling rate and poor stability in the preparation process of immunomagn

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
  • Preparation method for immunomagnetic microsphere, Sudan red detection kit containing immunomagnetic microsphere and detection method
  • Preparation method for immunomagnetic microsphere, Sudan red detection kit containing immunomagnetic microsphere and detection method
  • Preparation method for immunomagnetic microsphere, Sudan red detection kit containing immunomagnetic microsphere and detection method

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0029] The preparation process of immunomagnetic microspheres is usually as follows: the magnetic microspheres with chemical functional groups on the surface are activated by an activating agent in the activation buffer, and then the activated magnetic microspheres are placed in the coupling buffer, where they are mixed with The antibody or antigen is subjected to a coupling reaction, and after the coupling reaction is completed, the unreacted sites on the surface of the magnetic microspheres are blocked, and then the obtained immunomagnetic microspheres are used or dispersed in a storage solution for preservation. The activation of magnetic microspheres mostly uses MES (2-morpholineethanesulfonic acid) as a buffer, but the inventors have found that in the coupling reaction between some magnetic microspheres and antibodies or antigens, the coupling reaction obtained by using MES buffer The coupling rate is poor, and the obtained immunomagnetic microspheres have poor stability a...

Example Embodiment

[0066] Embodiment 1 is coated with the preparation of the immunomagnetic microsphere of Sudan red antibody

[0067] 1. Preparation of Reagents

[0068] 1) Activation / coupling buffer:

[0069] Preparation of 10mM PB buffer solution (pH6.0): ①Weigh 1.56g of sodium dihydrogen phosphate, and dilute to 1000mL with deionized water; ②Weigh 0.358g of disodium hydrogen phosphate, and dilute to 100mL with deionized water; Add ① reagent 17.54mL and ② reagent 2.46mL, then add Tween-20 and PVP to the obtained 10mMPB buffer, the volume percentage of Tween-20 is 0.05%, the mass percentage of PVP is 0.05%, and the validity period is 12 moon.

[0070] 2) Blocking solution:

[0071] Weigh 0.1 g of BSA solid and dissolve in 10 mL of coupling buffer. The reagent is a light yellow, clear and transparent liquid, and the prepared blocking solution can be stored at 2-8°C for 30 days.

[0072] 3) Storage solution:

[0073] Weigh 0.24 g of potassium dihydrogen phosphate, 3.58 g of disodium hydrog...

Example Embodiment

[0085] Embodiment 2 Formation of the Sudan Red Rapid Detection Kit Based on Magnetic Microspheres

[0086] Set up the Sudan Red rapid detection kit based on magnetic microspheres, so that it contains the following components:

[0087] (1) The immunomagnetic microspheres coated with the monoclonal Sudan red antibody No. 1 prepared in Example 1.

[0088] (2) Sudan red enzyme conjugate, which is HRP-labeled Sudan Ⅰ antigen (Sudan red antigen was purchased from Guangzhou Youkangduo Biotechnology Co., Ltd. (13mg / mL), HRP-labeled Sudan red antigen was obtained from Beijing Tianenze Gene Technology Co., Ltd. Limited (1mg / mL)).

[0089] (3) Sample diluent, the sample diluent is 0.01M PBS (phosphate) buffer containing 10% methanol and 0.1% Triton X-100, pH 7.4.

[0090] (4) Standard Sudan Red: Purchased from Beijing Beina Chuanglian Biotechnology Research Institute (100mg / bottle), prepare Sudan Red standard solutions with concentrations of 0, 0.2, 0.6, 1.8, 5.4ng / mL with sample diluent...

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 sizeaaaaaaaaaa
Particle sizeaaaaaaaaaa
Login to view more

Abstract

The invention provides a preparation method for an immunomagnetic microsphere. The method adopts a phosphate buffer containing 0.01%-0.1% of Tween-20 and 0.01%-0.1% of polyvinylpyrrolidone as the activating buffer and/or coupling buffer. The invention also discloses a Sudan red detection kit containing the immunomagnetic microsphere and a detection method using the kit to conduct detection. The immunomagnetic microsphere preparation method provided by the invention has obviously improved coupling rate, and the obtained immunomagnetic microsphere has good stability. The kit provided by the invention uses the Sudan red monoclonal antibody coated immunomagnetic microsphere, has high sensitivity, can be used for rapid screening of mass samples, can avoid the influence of the matrix in a to-be-detected sample, and has high accuracy. The detection method provided by the invention can play an important role in food safety detection, and has important social significance and broad market prospects.

Description

technical field [0001] The invention relates to the field of analysis and detection, in particular to a preparation method of immunomagnetic microspheres, a Sudan red detection kit containing immunomagnetic microspheres and a detection method. Background technique [0002] Sudan red is a kind of chemical dyeing agent, which is widely used in the color enhancement of solvents, oils, waxes and gasoline, as well as in the enhancement of shoes and floors. It is not a food additive, but because of its bright coloring, it is often illegally added to condiments such as paprika. It has an azo structure in its chemical composition. Due to the nature of this chemical structure, it is carcinogenic and has obvious toxic effects on the liver and kidney organs of the human body, so it is strictly prohibited from being added to food. [0003] At present, the detection methods of Sudan Red include high-performance liquid chromatography, liquid chromatography-mass spectrometry, etc. Althoug...

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
IPC IPC(8): G01N33/531G01N33/577
CPCG01N33/531G01N33/577
Inventor 王琳田静秒王雪万渝平周浩叶敏梁恒兴张敏
Owner BEIJING PURKINJE GENERAL INSTR
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