Orf virus (ORFV) protein ORFV059 monoclonal antibody hybridoma G3 and monoclonal antibody

A technology of ORFV059 and aphthous ulcer virus, which is applied in the field of cell engineering, can solve the problems of ineffective research on disease control and lack of specific antibodies, and achieve the effects of reduced application cost, high specificity and sensitivity

Inactive Publication Date: 2012-07-04
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, there have been several outbreaks of aphthous ulcers among sheep in Jilin Province and Gansu Province in...

Method used

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  • Orf virus (ORFV) protein ORFV059 monoclonal antibody hybridoma G3 and monoclonal antibody
  • Orf virus (ORFV) protein ORFV059 monoclonal antibody hybridoma G3 and monoclonal antibody
  • Orf virus (ORFV) protein ORFV059 monoclonal antibody hybridoma G3 and monoclonal antibody

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Embodiment 1

[0021] Example 1 Preparation and identification of the monoclonal antibody of the present invention

[0022] 1. Preparation of monoclonal antibody G3

[0023] 1) Preparation of recombinant ORFV059 antigen

[0024] Using the orf virus genome as a template, the full-length ORFV059 gene was obtained by PCR amplification, and the prokaryotic recombinant expression vector pET28a(+) / ORFV059 was constructed, which was expressed in Escherichia coli Escherichia coli Expressed in BL21, and purified by Ni Sepharose affinity chromatography purification column, the recombinant ORFV059 protein with a purity of more than 90% was obtained.

[0025] 2) Immunization of mice

[0026] 6-8 week old female BALB / c mice were immunized with purified recombinant antigen.

[0027] The first immunization: After mixing the recombinant antigen with an equal volume of Bentonite adjuvant, inject BALB / c mice intraperitoneally at 50 μg / 500 μL each;

[0028] The second immunization: After 2 weeks, take th...

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Abstract

The invention discloses an orf virus (ORFV) protein ORFV059 monoclonal antibody hybridoma G3 and a monoclonal antibody. The ORFV059 monoclonal antibody hybridoma G3 was collected in China General Microbiological Culture Collection Center (CGMCC) on November 4, 2011, wherein the collection number is CGMCC No.5425. The recombinant ORFV059 protein coats an enzyme-linked immuno sorbent assay (ELISA) plate; activity of a purified antibody is detected by an ELISA method; and the purified ORFV059 protein is subjected to Western-blot by the purified antibody, which proves that the antibody can identify the ORFV059 protein. The purified antibody is used for performing immunofluorescent staining on the ORFV059 and immunohistochemical staining on the orf tissue, which proves that the purified antibody can identify the natural ORFV059 protein. The monoclonal antibody secreted by the hybridoma cell lays basis for further development of ORFV059 function and a diagnosis reagent for developing the ORFV059 and provides a powerful tool for clinical diagnosis on the orf and confirmation of a therapeutic target.

Description

[0001] technical field [0002] The invention relates to the field of cell engineering, in particular to hybridoma cell line G3 and the monoclonal antibody of ORFV059 monoclonal antibody to the structural protein of oral ulcer virus. Background technique [0003] Ovine infectious impetigo (Orf), also known as ovine infectious impetigo, is an acute, contagious disease of goats and sheep caused by oral ulcer virus (ORFV). The lesion is characterized by the formation of erythema, papules, abscesses, ulcers and verrucous scabs on the skin and mucous membranes of the affected sheep. Lambs are very susceptible and often occur in groups. The disease occurs all over the world, and it is also common in the main sheep-raising areas of my country. It is one of the main diseases that harm sheep. The virus has strong resistance, and once the flock is infected, it is not easy to get rid of it. It can continue to harm the flock for many years and cause major economic losses to the animal...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/08G01N33/577
Inventor 罗树红郝文波李宏廖小青李明
Owner SOUTHERN MEDICAL UNIVERSITY
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