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Rabies virus monoclonal antibody and application thereof

A rabies virus and monoclonal antibody technology, which is applied to rabies virus monoclonal antibody and its application field, and achieves the effects of high detection sensitivity, strong application value and high neutralization titer

Inactive Publication Date: 2014-07-30
北京凯思百奥科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Because rabies virus glycoprotein is difficult to express and purify in vitro, there are relatively few studies on monoclonal antibodies against rabies virus glycoprotein or monoclonal antibodies with neutralizing rabies virus activity

Method used

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  • Rabies virus monoclonal antibody and application thereof
  • Rabies virus monoclonal antibody and application thereof
  • Rabies virus monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] 1. Establishment of hybridoma cell lines

[0024] (1) Experimental materials

[0025] 1. Immunogen: In this case, the inactivated virus of the CVS strain of rabies virus was used as the immunogen.

[0026] 2. Culture medium: DMEM medium was purchased from GbiCo; HAT and HT selection medium.

[0027] 3. Experimental animals: Balb / c mice, 8-12 weeks old, female, SPF grade animal culture.

[0028] 4. Other materials: Freund's complete adjuvant and Freund's incomplete adjuvant were purchased from Sigma Company; PEG4000 was purchased from Fluka Company; HRP-goat anti-mouse IgG antibody was purchased from Sigma Company; other reagents were domestic analytically pure products.

[0029] (2) Establishment of hybridoma cell lines

[0030] 1. Animal immunity

[0031] 1) Basic immunization: the antigen was mixed with Freund's complete adjuvant in equal volumes and fully emulsified, and injected subcutaneously in points, each injection volume of each Balb / c mouse was 100 μg.

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Abstract

The invention relates to a rabies virus monoclonal antibody with neutralization activity and capability of specifically recognizing rabies virus glycoprotein G. The monoclonal antibody is authenticated to be IgG1-subtype. According to the rabies virus monoclonal antibody disclosed by the invention, rabies virus inactivation totivirus is taken as an immunogen for immunizing a mouse, an antibody with neutralization activity is screened, the neutralization titer is high, the secretion of hybridoma for the antibody is stable, and the antibody is easy to store; and an in-vitro preparation link for glycoprotein is avoided, and innovativeness is achieved. A colloidal gold test paper card or test paper strip prepared from the antibody is high in test sensitivity and capable of achieving 0.03 IU / mL, capable of being used for test on human rabies virus or quality control in a vaccine production process, and high in application value.

Description

technical field [0001] The invention relates to a rabies virus monoclonal antibody and its preparation method and application. Background technique [0002] Rabies is a deadly zoonotic disease caused by Rabies Virus (RV) infection. Once the infected person becomes ill, almost 100% of them die. Rabies virus belongs to the family Rhabdoviridae and the genus Lyssavirus. The genome is a single-stranded negative-strand RNA without segments, with a molecular weight of about 4.6×106D and a size of about 12kb. It encodes five viral structural proteins, namely nucleoprotein (N), phosphoprotein (P), matrix protein (M), Glycoprotein (G), RNA polymerase (L). Among them, rabies virus glycoprotein G is a transmembrane protein, which constitutes the surface protrusions of the virus and belongs to protective antigens. It can not only induce the body to produce protective neutralizing antibodies, but also stimulate the proliferation of T lymphocytes. [0003] All warm-blooded animals are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/569
CPCG01N33/68G01N33/531G01N33/56983G01N33/577G01N2333/145
Inventor 邢江峰
Owner 北京凯思百奥科技发展有限公司
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