Application of a da monoclonal antibody immunoaffinity column in the determination of domoic acid toxoids

A monoclonal antibody and immunoaffinity technology, applied in the field of immunoaffinity columns, can solve the problems of shortening analysis time and unsatisfactory sample purification, and achieve the effect of shortening analysis time, solving unsatisfactory sample purification and high performance

Active Publication Date: 2021-06-22
MARINE FISHERIES RES INST OF ZHEJIANG
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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

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Examples

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Effect test

Embodiment

[0033] (a) Synthetic immunogens

[0034] Add 10mg bovine serum albumin, 2mL 0.05M, pH7.4 sodium acetate buffer, 1mg domoic acid, 60μL formaldehyde solution with a mass concentration of 37% to the small flask, mix well, stir at 37°C for 48h, 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 reagents

[0036] Add 10mg of chicken ovalbumin, 2mL of 0.05M, pH7.4 sodium acetate buffer, 1mg of domoic acid, and 60μL of formaldehyde solution with a mass concentration of 37% in a small flask, mix well, and stir at 37°C for 48h. 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) Preparation and purification of monoclonal antibodies

[0038] Five female Balb / c mice aged 6-8 weeks were injected subcutaneously with DA-BSA at multiple points on the back, 20 μg / mouse, and the boo...

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Abstract

The present invention relates to an immunoaffinity column, in particular to a method for preparing a DA monoclonal antibody immunoaffinity column. A DA monoclonal antibody with high performance and strong recognition is prepared by screening out antibodies specific to DA toxin. Cloned antibody immunoaffinity column, the obtained DA monoclonal antibody immunoaffinity column can be applied to the analysis and determination of DA toxin in different marine organisms, which can effectively solve the problem of unsatisfactory sample purification in pretreatment, reduce sample matrix interference, and shorten the analysis time. time, improve the recovery rate of the method, and fill the gap of immunoaffinity technology in the field of DA toxin detection. The invention also provides an application of the DA monoclonal antibody immunoaffinity column.

Description

technical field [0001] The invention relates to an immunoaffinity column, in particular to the application of a DA monoclonal antibody immunoaffinity column in the determination of domoic acid toxoids. 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. ...

Claims

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Application Information

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Patent Type & Authority Patents(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
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