Monoclonal antibody of toxoplasma gondii resistant MIC3 protein and application monoclonal antibody

A monoclonal antibody, Toxoplasma gondii technology, applied in immunoglobulins, biochemical equipment and methods, instruments, etc., can solve the problems of high experimental conditions, specificity needs to be improved, difficult to popularize and apply, etc., and achieve the antibody purification process. Simple, specific, and low-cost effects

Active Publication Date: 2013-10-02
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Etiological examination, such as microscopic examination of parasites from body fluids and tissues, animal inoculation, etc., can confirm the diagnosis by finding parasites, but the detection rate is low and time-consuming; serological examinations include staining test (DT), enzyme-linked immunosorbent assay ( ELISA), indirect hemagglutination assay (IHA), immunoblotting and other methods have high sensitivity, but specificity needs to be improved; genetic examination, such as DNA probe technology, polymerase chain reaction and other methods, have high sensitivity and specificity However, the operation is complicated and the experimental conditions are relatively high; the IgM and IgG detection kits are expensive, and it is difficult to promote the application and epidemiological census

Method used

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  • Monoclonal antibody of toxoplasma gondii resistant MIC3 protein and application monoclonal antibody
  • Monoclonal antibody of toxoplasma gondii resistant MIC3 protein and application monoclonal antibody
  • Monoclonal antibody of toxoplasma gondii resistant MIC3 protein and application monoclonal antibody

Examples

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Embodiment 1

[0032] Example 1 Preparation of monoclonal antibody against Toxoplasma gondii MIC3 protein

[0033] Prepare anti-toxoplasma MIC3 protein monoclonal antibody according to the following method: use E. coli expression system to prepare recombinant MIC3 protein; use recombinant MIC3 protein as antigen to immunize mice, prepare fusion hybridoma cells, and obtain stable secretion of anti-toxoplasma by limiting dilution method The hybridoma cell line D3 is a monoclonal antibody against worm MIC3 protein.

[0034] 1 Preparation of recombinant MIC3 protein

[0035] 1. Construction of positive recombinant plasmid pET-32a(+)-MIC3

[0036] The nucleotide sequence shown in SEQ ID NO:1 (synthesized by Nanjing KingScript Biotechnology Co., Ltd.) and the pET-32a (+) expression vector were respectively used with restriction endonuclease H ind III and SacⅠ double digestion, the fragments were recovered by 1.5% agarose gel electrophoresis and ligated, and the ligated products were transforme...

Embodiment 2

[0066] Example 2 Preparation and Properties of Toxoplasma Gondii Antigen Colloidal Gold Immunochromatography Detection Test Strip

[0067] (1) Preparation of 20-30nm colloidal gold particles

[0068] The gold solution was prepared by trisodium citrate reduction method. Get 100mL of 0.01% chloroauric acid aqueous solution and heat to boiling, accurately add 1.8mL of 1% trisodium citrate aqueous solution under stirring, continue to boil for 10min, add distilled water after cooling and make it return to the original volume, measure the absorption peak wavelength of gold solution, avoid Store at 4°C. After scanning and observing with a UV-visible spectrophotometer, the measured absorption curve is the absorption peak at 521nm, and the peak scanning result is D 521nm =0.857, the corresponding particle diameter is 20-30nm.

[0069] (2) Preparation of gold standard pad

[0070] Take 20 mL of colloidal gold solution (particle diameter: 20-30 nm) with pH 7.5, and slowly add the...

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Abstract

The invention provides a monoclonal antibody of a toxoplasma gondii resistant MIC3 protein and an application of the monoclonal antibody, and relates to the technical field of biology. The preservation number of a hybridoma cell strain D3 of secreting the monoclonal antibody of the toxoplasma gondii resistant MIC3 protein is CCTCCC (China Center For Type Culture Collection) 201383. The invention also discloses the monoclonal antibody of the toxoplasma gondii resistant MIC3 protein secreted by the hybridoma cell strain D3, and an application of the monoclonal antibody of the toxoplasma gondii resistant MIC3 protein in preparation of a kit and a colloidal gold test strip for detecting the toxoplasma gondii. The monoclonal antibody of the toxoplasma gondii resistant MIC3 protein can be stably and efficiently secreted by the hybridoma cell strain D3; and the monoclonal antibody has high titer and good specificity on the toxoplasma gondii MIC 3 protein. Therefore, the monoclonal antibody can be used for preparing the kit and the colloidal gold test strip for detecting the toxoplasma gondii. The monoclonal antibody is simple in preparation method, simple in anti-body purification process, high in efficiency and low in cost.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an anti-toxoplasma MIC3 protein monoclonal antibody and application thereof. Background technique [0002] Toxoplasmosis is a zoonotic parasitic disease that is distributed worldwide and seriously endangers human health and the development of animal husbandry. Humans can become infected with toxoplasmosis by eating undercooked pork that contains Toxoplasma cysts. Toxoplasmosis can cause miscarriage, premature birth, teratogenicity, and stillbirth in pregnant women. [0003] Toxoplasma detection methods have a variety of methods. Etiological examination, such as microscopic examination of parasites from body fluids and tissues, animal inoculation, etc., can confirm the diagnosis by finding parasites, but the detection rate is low and time-consuming; serological examinations include staining test (DT), enzyme-linked immunosorbent assay ( ELISA), indirect hemagglutination assay (IHA)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/20G01N33/577G01N33/569G01N33/558
Inventor 白昀邵国青刘茂军冯志新王海燕白方方熊祺琰王占伟甘源刘冬霞
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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