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Feline calicivirus monoclonal antibody and application thereof

A monoclonal antibody and feline calicivirus technology, applied in antiviral immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems of inability to accurately detect strong FCV strains, and achieve good reactivity and high sensitivity , the effect of high accuracy

Active Publication Date: 2019-09-24
LUOYANG PULIKE WANTAI BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing products cannot accurately detect FCV virulent strains. In order to face this severe situation, it is urgent to establish a fast, accurate, simple and broad-spectrum diagnostic method for FCV.

Method used

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  • Feline calicivirus monoclonal antibody and application thereof
  • Feline calicivirus monoclonal antibody and application thereof
  • Feline calicivirus monoclonal antibody and application thereof

Examples

Experimental program
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Embodiment approach

[0031] As an embodiment of the present invention, the antibody is a monoclonal antibody or a genetically engineered antibody; wherein, the genetically engineered antibody includes a single-chain antibody, a chimeric monoclonal antibody, a modified monoclonal antibody, or a fragment of the antibody ; said antibody or a fragment of said antibody still retains the ability to specifically bind feline calicivirus.

[0032] As an embodiment of the present invention, the antibody is monoclonal antibody 5D9, the amino acid sequence of the heavy chain variable region of the monoclonal antibody 5D9 is the amino acid sequence encoded by SEQ.ID No.1, and the amino acid sequence of the light chain variable region is The sequence is the amino acid sequence encoded by SEQ.ID No.2.

[0033] The monoclonal antibody 5D9 is a feline calicivirus monoclonal antibody, and is further a monoclonal antibody to the feline calicivirus Cap protein; the amino acid sequence of the heavy chain variable regi...

Embodiment 1

[0067] Isolation and identification of embodiment 1 feline calicivirus

[0068] Collect cats with clinical symptoms such as oral ulcers, rhinitis, conjunctivitis, chronic gastroenteritis, acute arthritis, anorexia, lethargy and even death due to these clinical symptoms (from pets in Henan, Shandong, Guangdong, Hubei, Sichuan and other provinces) 50 eye, nose or nasopharyngeal swabs from the hospital) were detected with the feline calicivirus RT-PCR detection kit. Except for 10 negative, the remaining 40 were positive, and 30 of them were related to feline herpes virus 1 Type mixed infection, or mixed infection with feline panleukopenia virus, or mixed infection with three viruses, only 10 copies were infected with FCV single virus, the corresponding disease material was inoculated into FK81 cells, and the corresponding virus was obtained by blind passage for 3 generations, and harvested Virus culture fluid and its structural proteins were sequenced. The result: the structural ...

Embodiment 2

[0069] Preparation, purification, identification and inspection of embodiment 2 feline calicivirus monoclonal antibody

[0070] 2.1 Preparation and purification of feline calicivirus monoclonal antibody

[0071] 10 FCV strains (10 strains) prepared in Example 1 5.0 TCID 50 / ml) concentrated with PEG20000 as the immunogen respectively, emulsified with Freund's complete adjuvant for the first immunization, and after emulsifying with Freund's incomplete adjuvant for the second and third immunizations, immunize 6-8 week-old females at 0.4ml / cause Balb / C mice, 10 groups in total. First use the IFA antigen plates prepared by 10 FCV strains to detect the IFA titer of the mouse serum after the three immunizations of the respective strains as the immunogen, and select the mouse serum with the highest IFA titer (see mice 1-10 in Table 1) These 10 FCV strains were then used for IFA evaluation, and the results are shown in Table 1.

[0072] Indirect immunofluorescence IFA: 10 strains ...

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Abstract

The invention provides a variable region sequence of a monoclonal antibody specifically bound to a Feline calicivirus, an antibody specifically bound to a Feline calicivirus and a hybridoma cell secreting the antibody. The variable region sequence can be specially bound to the Feline caliciviruses, and can be combined with various Feline caliciviruses. A dual-antibody sandwich detection reagent kit prepared by the antibody can be used for detecting varied FCV toxin strains clinically, the problems that through detection in the prior art, the broad spectrum of the FCV toxin strains is poor, and the sensitivity is low, are solved, the phenomena of being undetected and false-negative are avoided, particularly the accuracy of detecting the targets of varied types of asymptomatic animals is higher, and the variable region sequence has the advantages of being quick, simple and accurate.

Description

technical field [0001] The invention relates to two kinds of feline calicivirus monoclonal antibodies, a double-antibody sandwich detection kit containing monoclonal antibody pairs and an application thereof, belonging to the field of biotechnology. Background technique [0002] Feline calicivirus (FCV) belongs to the family Caliciviridae and can cause feline (mainly cats) upper respiratory tract infection, oral vesicular disease and chronic gastroenteritis, as well as feline pneumonia and lameness syndrome and other diseases. The incidence of disease caused by FCV is extremely high, and it is distributed worldwide, and it is very easy to infect kittens within one year old, resulting in the death of kittens. FCV has a wide range of transmission routes, and the main source of infection is sick cats and infected cats. In the acute stage, sick cats can excrete a large amount of virus with secretions and excretions, and directly infect susceptible cats; after treatment, infecte...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N5/20G01N33/577G01N33/569C12R1/91
CPCC07K16/10G01N33/577G01N33/56983C07K2317/56C07K2317/622
Inventor 田克恭王莹王景云
Owner LUOYANG PULIKE WANTAI BIOTECH
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