Monoclonal antibody specially bound to TGEV (transmissible gastroenteritis of swine virus), pharmaceutical composition, kit and applications of monoclonal antibody, pharmaceutical composition and kit

A monoclonal antibody and infectious technology, applied in the direction of antiviral agents, antibodies, antiviral immunoglobulin, etc., can solve the problems of complication of pig disease in epidemic areas, aggravation of pig disease, easy mixed infection, etc.

Active Publication Date: 2019-01-18
LUOYANG PULIKE WANTAI BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In clinical diagnosis, the two are very easy to mix infection, which will complicate the disease of pigs in the epidemic area, which will not only affect the accurate and effective detection results and guiding countermeasures given by the inspectors, but also aggravate the incidence of pigs , resulting in greater losses
[0004] At present, the methods for detecting porcine transmissible gastroenteritis virus include virus isolation and identification and serum examination, but it takes too long; although the PCR method is sensitive, it requires expensive instruments and professional operators, and it cannot be detected in tissues and cells. Carry out clear positioning, cannot track the dynamic distribution of pathogens in the body, and semi-quantitative detection

Method used

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  • Monoclonal antibody specially bound to TGEV (transmissible gastroenteritis of swine virus), pharmaceutical composition, kit and applications of monoclonal antibody, pharmaceutical composition and kit
  • Monoclonal antibody specially bound to TGEV (transmissible gastroenteritis of swine virus), pharmaceutical composition, kit and applications of monoclonal antibody, pharmaceutical composition and kit
  • Monoclonal antibody specially bound to TGEV (transmissible gastroenteritis of swine virus), pharmaceutical composition, kit and applications of monoclonal antibody, pharmaceutical composition and kit

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Experimental program
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Effect test

Embodiment approach

[0021] As an embodiment of the present invention, the antibody is a monoclonal antibody and a genetically engineered antibody; wherein, the genetically engineered antibody includes a single-chain antibody, a chimeric monoclonal antibody, a modified monoclonal antibody, and a porcine monoclonal antibody or a fragment of said antibody; said antibody or said fragment of said antibody still retains the ability to specifically bind porcine transmissible gastroenteritis virus.

[0022] The term "transmissible gastroenteritis of swinevirus" (transmissible gastroenteritis of swinevirus, TGEV) belongs to the Coronaviridae (Coronaviridae) genus (Coronavirus), is a single-stranded positive-sense RNA virus, and is easy to cause vomiting, diarrhea, dehydration, high A porcine transmissible gastroenteritis disease characterized by death.

[0023] The term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, ie, the individual antibo...

Embodiment 1

[0081] Example 1 Preparation, Purification, Identification and Application of Porcine Transmissible Gastroenteritis Virus Monoclonal Antibody

[0082] 1.1 Preparation and purification of monoclonal antibodies 3D2 and 4B4

[0083] Cultivate porcine transmissible gastroenteritis virus Hua strain (purchased from Wuhan University China Type Culture Collection Center, preservation number is CCTCC-V200609), according to Su Xue et al. (Su Xue. Porcine transmissible gastroenteritis virus monoclonal antibody Preparation and Identification. Inner Mongolia Agricultural University. Master Thesis) method to purify virus and determine OD 280 , the calculated protein concentration after purification is 1.67mg / ml; it is emulsified with Freund's adjuvant, and the immunogenic composition is prepared according to the final content of 100μg / mouse, and the mice are immunized every 2 weeks. Blood was collected from the mice, and the cells infected with Sinovirus strain were used as indirect immuno...

Embodiment 2

[0128] Preparation and application of embodiment 2 kit

[0129] 2.1 Pairing studies of monoclonal antibodies

[0130] Dilute TGEV Hua strain and HB08 strain to 10 5.0 TCID 50 / ml, different matching modes were tested, and the results were the same, see Table 4:

[0131] Table 4 Matching results of monoclonal antibodies

[0132]

[0133] Note: "+" means positive, "-" means negative.

[0134] Results: The detection of porcine transmissible gastroenteritis virus liquid by labeled monoclonal antibody 4B4 and immobilized monoclonal antibody 3D2 was negative, and the results of other combinations were positive. Therefore, the combination of immobilized monoclonal antibody 4B4 and labeled monoclonal antibody 3D2 was selected for kit preparation.

[0135] 2.2 Preparation and application of TGEV colloidal gold test strip

[0136] 2.2.1 Preparation

[0137] HAuC1 4 First prepare a 0.01% aqueous solution, take 100ml and heat it to boiling, and accurately add 1.0ml of 1% trisod...

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Abstract

The invention provides a monoclonal antibody specially bound to TGEV (transmissible gastroenteritis of swine virus) and a kit prepared from the monoclonal antibody. The TGEV can be determined rapidlyand accurately, and strains can be widely determined. Pharmaceutical composition prepared from the antibody can be used for broad-spectrum prevention and treatment of different TGEVs.

Description

technical field [0001] The invention relates to a monoclonal antibody capable of specifically binding porcine transmissible gastroenteritis virus, and a pharmaceutical composition, kit and application prepared by using the monoclonal antibody, belonging to the field of biomedicine. Background technique [0002] Transmissible gastroenteritis of swine (TGE) is a highly contagious digestive tract infectious disease caused by transmissible gastroenteritis of swine virus (TGEV), which mainly causes vomiting, Symptoms such as watery diarrhea and dehydration can cause the death of piglets in severe cases, and the mortality rate can reach 50%-100%. In intensive breeding areas, TGEV is considered to be one of the main culprits causing morbidity and mortality in piglets. The disease has been reported in many countries such as Europe, North America and Asia. In recent years, porcine transmissible gastroenteritis has occurred and spread in some areas of our country from time to time, w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10G01N33/577G01N33/569G01N33/558A61K39/42A61P31/14
CPCA61K2039/505C07K16/10C07K2317/56G01N33/558G01N33/56983G01N33/577G01N2333/17
Inventor 田克恭张云静
Owner LUOYANG PULIKE WANTAI BIOTECH
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