Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Myoglobin antibody-enzyme marker as well as preparation and application thereof

A myoglobin and enzyme labeling technology, applied in the field of antibody-enzyme markers and their preparation and application, can solve the problems of unfavorable large-scale production, difficult storage, difficult operation, etc., and achieve easy operation, less by-products, and background low value effect

Active Publication Date: 2021-08-27
NINGBO RUI BIO TECH
View PDF10 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The hydroxyl group of the sugar-containing part of ALP is oxidized into aldehyde groups by sodium periodate, and combined with antibodies under certain conditions to form a Schiff base, and then the redundant aldehyde groups are reduced by sodium borohydride. This method is difficult to operate and expensive. Sodium iodate is a strong oxidant and is not easy to store. Sodium borohydride is a strong reducing agent, which will reduce water to hydrogen in the reduction step, which is not conducive to large-scale production

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
  • Myoglobin antibody-enzyme marker as well as preparation and application thereof
  • Myoglobin antibody-enzyme marker as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation method of myoglobin antibody-enzyme marker may specifically comprise the following steps:

[0040] (1) Preparation and purification of maleimide-modified myoglobin antibody MB II (MB II-MHS) Weigh 500ug of myoglobin antibody MB II and dissolve it in 5mL of 0.1M PBS with pH=7.5;

[0041] Add MHS with a molar ratio of about 10:1 [floating range (10±1):1] to myoglobin antibody MB II, and react in the dark at room temperature for 2±0.2 hours to obtain an incubation solution;

[0042] Take out the incubation solution, put it into a dialysis bag, and dialyze in a 5mM PBS solution in a refrigerator at 4°C for 12±1.2 hours, changing the solution 3 to 4 times during the period;

[0043] Collect the dialyzed incubation solution and store it at 2-8°C for later use;

[0044] (2) Preparation and purification of thiol-modified enzyme label (ALP-SATA)

[0045] Weigh 500ug of alkaline phosphatase and dissolve it in 5mL of 0.1M, pH=7.5 PBS;

[0046] Add SATA with a mo...

Embodiment 2

[0053] The only difference from Example 1 is that in the step (1), the molar ratio of 6-(maleimido) hexanoic acid succinimide ester to myoglobin antibody is 8:1, and other Example 1 remains the same.

Embodiment 3

[0055] The only difference from Example 1 is that in the step (1), the molar ratio of 6-(maleimido) caproic acid succinimide ester to myoglobin antibody is 13:1, and other Example 1 remains the same.

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

No PUM Login to View More

Abstract

The invention relates to the technical field of enzyme immunity, in particular to a myoglobin antibody-enzyme marker as well as preparation and application thereof. The myoglobin antibody-enzyme marker is obtained by crosslinking a maleimide modified myoglobin antibody MB II and a sulfydryl modified enzyme marker, wherein the maleimide modified myoglobin antibody MB II is prepared by taking 6-(maleimide) hexanoic acid succinimide ester as a cross-linking agent and cross-linking the 6-(maleimide) hexanoic acid succinimide ester with a myoglobin antibody; and the sulfydryl modified enzyme marker is prepared by taking N-succinimide S-acetylthioacetate as a cross-linking agent, cross-linking the cross-linking agent with alkaline phosphatase and then activating the cross-linking agent with hydroxylamine hydrochloride. When the myoglobin antibody-enzyme marker is used for enzymatic chemiluminescence immunoassay after being diluted by a diluent, compared with a common antibody-enzyme marker, the background value of the myoglobin antibody-enzyme marker is 3-5 times lower, and the ratio of a high-value signal to a low-value signal is 3 times or above higher.

Description

technical field [0001] The invention relates to the technical field of enzyme immunity, in particular to an antibody-enzyme marker and its preparation and application. Background technique [0002] In the detection of enzyme-catalyzed chemiluminescence immunoassay, the antigen or antibody enzyme marker plays an important role in the sensitivity and specificity of immunoassay, and is the key reagent in the detection of enzyme-catalyzed chemiluminescence immunoassay. Alkaline phosphatase (alkaline phosphatase, ALP) is an enzyme that is widely used in enzymatic chemiluminescent immunodiagnostic reagents. Combining it with antibodies through certain technical means is a very important step in the development of enzymatic chemiluminescent immunodiagnostic reagents. . In general, there are mainly the following methods for cross-linking alkaline phosphatase and antibody: [0003] 1. Glutaraldehyde method [0004] The methods for cross-linking antibodies with glutaraldehyde as a ...

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): G01N33/72G01N33/535
CPCG01N33/72G01N33/535G01N2333/805
Inventor 艾杨洋陈媛张闻
Owner NINGBO RUI BIO TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com