Preparation method and application of DA monoclonal antibody immunoaffinity column

A monoclonal antibody and immunoaffinity technology, applied in the field of immunoaffinity columns, can solve the problems of unsatisfactory sample purification, shorten analysis time, etc., and achieve the effect of solving unsatisfactory sample purification, shortening analysis time, and improving recovery rate

Active Publication Date: 2019-03-19
MARINE FISHERIES RES INST OF ZHEJIANG
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for preparing a DA monoclonal antibody immunoaffinity column. By screening out antibodies specific to DA toxin, the prepared immunoaffinity column has the characteristics of high performance and strong recognition. It is applied to the analysis and determination of DA toxin in different marine organisms, effectively solves the problem of unsatisfactory sample purification in pretreatment, reduces sample matrix interference, shortens analysis time, improves method recovery rate, and fills the gap of immunoaffinity technology in the field of DA toxin detection. blank

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0033] (a) Synthetic immunogens

[0034] Add 10 mg of bovine serum albumin, 2 mL of 0.05 M, pH7.4 sodium acetate buffer, 1 mg of domoic acid, and 60 μL of formaldehyde solution with a mass concentration of 37% in a small flask, mix well, and stir at 37 ° C for 48 h. Then use 0.1M, pH7.3 PBS buffer solution, dialyze at 4°C for 3 days, change the solution twice a day, and obtain the immunogen DA-BSA;

[0035] (b) Synthetic detection agent

[0036] Add 10 mg of chicken ovalbumin, 2 mL of 0.05 M, pH7.4 sodium acetate buffer, 1 mg of domoic acid, and 60 μL of formaldehyde solution with a mass concentration of 37% in a small flask, mix well, and stir at 37 ° C for 48 h. Then use 0.1M, pH7.3 PBS buffer solution, dialyze at 4°C for 3 days, change the solution twice a day, and detect the original DA-OVA;

[0037] (c) Monoclonal antibody preparation and purification

[0038] Five female Balb / c mice at the age of 6-8 weeks were subcutaneously injected with DA-BSA at multiple points on...

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
collision energyaaaaaaaaaa
Login to view more

Abstract

The invention relates to an immunoaffinity column and particularly relates to a preparation method of a DA monoclonal antibody immunoaffinity column. By screening an antibody with the specificity to DA toxin, the high-perforamnce and strong-recognition capacity DA monoclonal antibody immunoaffinity column is prepared and can be applied to the assay determination of DA toxin in different marine organisms, so that the problem that the purification of a sample in the pretreatment is not ideal is effectively solved, the matrix interference of the sample is reduced, the analysis time is shortened,the recovery rate of the method is increased, and the blank of an immunoaffinity technique in the detection field of DA toxin is filled up. The invention further provides application of the DA monoclonal antibody immunoaffinity column.

Description

technical field [0001] The invention relates to an immunoaffinity column, in particular to a preparation method and application of a DA monoclonal antibody immunoaffinity column. Background technique [0002] Amnestic shellfish poisoning (ASP) is a toxin secreted by red tide diatoms, and its main active ingredient, domoic acid (DA), is a neurogenic amino acid toxoid. The toxin was first discovered in 1987 in a poisoning death caused by eating blue mussels on the east coast of Prince Edward Island, Canada. Poisoning mainly manifests as abdominal pain, diarrhea, and vomiting, accompanied by severe headache, memory loss, confusion, coma and other symptoms. The study found that DA was mainly produced by Pseudonitzia multicolumns. When the content of domoic acid in shellfish tissue reaches 40mg / kg, it can cause poisoning to the eater, and when it is 150mg / kg, there is a risk of death. The maximum limit that humans can tolerate through eating is 20mg / kg. g Shellfish, Europe and...

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): G01N30/88G01N30/06G01N33/532G01N33/577
CPCG01N30/06G01N30/88G01N33/532G01N33/577G01N2030/8813
Inventor 张小军陈思严忠雍
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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