Celine parvovirus neutralizing monoclonal antibody and application thereof

A monoclonal antibody and feline parvovirus technology, which is applied in the field of bioengineering, can solve the problem that the neutralizing antibody treatment method is not widely used in clinical cases, etc., and achieves the effects of high neutralizing activity, convenient operation and simple preparation.

Active Publication Date: 2022-07-29
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Effective neutralizing antibody therapy has not been widely used in clinical cases

Method used

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  • Celine parvovirus neutralizing monoclonal antibody and application thereof
  • Celine parvovirus neutralizing monoclonal antibody and application thereof
  • Celine parvovirus neutralizing monoclonal antibody and application thereof

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Embodiment 1

[0031] 1. Purification of Feline Parvovirus

[0032] (1) FPV-susceptible cells CRFK were cultured to a monolayer in a 6-well culture plate, feline parvovirus was inoculated into CRFK, adsorbed in a 37°C incubator for 1-2 hours, then discarded the culture medium, and washed three times with PBS CRFK cells. The DMEM maintenance solution containing 2% fetal bovine serum was added to continue the culture at 37°C. Because cytopathic changes were difficult to observe, the virus proliferation was determined by indirect immunofluorescence at different time points. The results are as follows: figure 1 As shown, the highest viral load was found at 60 h, after which the cells began to shed. Therefore, at 60h, the first-generation culture was taken out, freeze-thawed 3 times between -80°C and normal temperature, centrifuged at 8000rpm for 15min at 4°C, and the supernatant was taken and dispensed into sterile EP tubes, which was the original virus. , recorded as P0 on behalf of the poiso...

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Abstract

The invention discloses a cat parvovirus neutralizing monoclonal antibody and application thereof, and relates to the technical field of bioengineering. The invention provides a hybridoma cell strain which is preserved in China Center for Type Culture Collection on May 6, 2022, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: C2022111. The invention also provides a cat parvovirus neutralizing monoclonal antibody which is secreted by the hybridoma cell, and the specific sequences of the cat parvovirus neutralizing monoclonal antibody are shown as SEQ ID NO: 1 and SEQ ID NO: 2. The hybridoma cell can secrete and generate the cat parvovirus neutralizing monoclonal antibody, and the monoclonal antibody is proved to have very high neutralizing activity through in-vitro cellular level neutralization tests and animal protective experiments, can specifically treat FPV, is simple to prepare and convenient to operate, and has great significance for preventing and treating the FPV.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a feline parvovirus neutralizing monoclonal antibody and its application. Background technique [0002] Feline panleukopenia virus (FPV), formerly known as feline distemper virus, also known as feline panleukopenia virus, feline panleukopenia virus, the clinical manifestations of infection are sudden high fever, intractable vomiting, diarrhea, dehydration The virus is highly contagious, with the highest morbidity and mortality in young animals. For cats, cats of all ages can be infected, and it mainly infects kittens under 1 year old. Unvaccinated kittens at 3-5 months of age are the most susceptible, with an infection rate of 70% and a case fatality rate of 50%-60%. The virus has a wide host range, including domestic and wild cats and some wild canids, such as raccoons and foxes, except dogs. It is currently the most widespread and most pathogenic virus of carnivore parv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/08A61K39/42A61P31/20
CPCC07K16/081A61P31/20C07K2317/76A61K2039/505
Inventor 彭贵青焦宇洲沈洲杨梦芳方祾颖史佳乐许谦
Owner HUAZHONG AGRI UNIV
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