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Anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10, and monoclonal antibody

A technology of H3N2-CIV-NP-2D10, monoclonal antibody, applied in the field of immunology, can solve problems such as influenza pandemic, achieve the effect of high specificity and sensitivity, and high secretion yield

Active Publication Date: 2017-02-08
GUANGZHOU YOUDI BIOTECH CO LTD
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Problems solved by technology

Recent studies have found that the HA gene of H3N2 subtype canine influenza virus may be introduced into human influenza virus through reassortment, and the introduction of new HA gene may lead to human-to-human influenza pandemic

Method used

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  • Anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10, and monoclonal antibody
  • Anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10, and monoclonal antibody
  • Anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10, and monoclonal antibody

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Embodiment Construction

[0019] The present invention will be further described below in conjunction with specific experiments, but not limited thereto.

[0020] Establishment of H3N2-CIV-NP-2D10 hybridoma cell line and preparation and identification of anti-H3N2 subtype canine influenza virus NP monoclonal antibody

[0021] 1. Immunization of mice

[0022] The inactivated H3N2 subtype canine influenza virus was mixed with an equal volume of complete Freund's adjuvant, and then injected subcutaneously into the back of BALB / C mice at multiple points in the amount of 50 μg / 500 μl each (Experimental Animal Center of Southern Medical University , 6-8 weeks old female, 3) for the first immunization; 2 weeks later, the adjuvant was replaced with Freund's incomplete adjuvant for the second routine immunization, the immunization method and dose were the same as the first immunization; a small amount was collected after three routine immunizations Tail vein blood, serum titer was detected by indirect ELISA. ...

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Abstract

The invention discloses an anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10, and a monoclonal antibody. The above hybridoma cell strain is preserved in China Center for Type Culture Collection on August 12, 2016 with the preservation number of CCTCC NO:C2016155. The hybridoma cell strain obtained in the invention has high secretion output, and the monoclonal antibody prepared for an H3N2 subtype canine influenza virus NP has high specificity and sensitivity, can bind to a denaturated NP protein in ELISA and Western-blot detection, can bind a natural NP protein with a high-grade structure in immunofluorescence detection, can provide scientific bases for researches of the structure functions of the NP protein and development of various subtype in-vitro diagnosis reagents, and provides experiment data support for the feasibility of the monoclonal antibody used as a disease marker.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to an anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10 and the monoclonal antibody. Background technique [0002] Influenza A virus is a common influenza virus. Influenza A virus is most likely to mutate. Influenza A virus includes multiple subtypes. Multiple subtypes of influenza viruses have been found to infect dogs, including H1, H3, H5, and H9 subtypes. Among them, H3N8 and H3N2 subtype influenza viruses can circulate in dogs, mainly causing respiratory symptoms. The emergence of canine influenza not only threatens the health of dogs, but also poses a threat to public health. Recent studies have found that the HA gene of H3N2 subtype canine influenza virus may be introduced into human influenza virus through reassortment, and the introduction of new HA gene may lead to human-to-human influenza pandemic. [0003] Canine influenza ...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/10G01N33/569G01N33/577
CPCC07K16/1018C07K2317/515
Inventor 张立成王燕李来庆梁焕坤陈翠翠王珊霞李康燕刘晓柱李佳丽刘细潘钟树海
Owner GUANGZHOU YOUDI BIOTECH CO LTD
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