Canine influenza virus monoclonal antibody hybridoma cell strain F112 and application thereof

Abstract

The invention relates to canine influenza virus monoclonal antibody hybridoma cell strain F112 and belongs to the technical field of biology. Immune antigens are prepared by using canine influenza virus epidemic strain A / Canine / Nanjing / 11 / 2012 (H3N2) through lymphocyte hybridoma technique, the efficient hybridoma cell strain F112 is screened out from the antigens, and the canine influenza virus monoclonal antibody hybridoma cell strain F112 has hemagglutination inhibition titer of 212 and neutralizing titer of 105 for canine influenza virus. A canine influenza preventive therapeutic agent prepared by using F112 monoclonal antibody is used for preventing and treating canine influenza, under the effective rate of 100%. The efficient F112 monoclonal antibody is applicable to the detection and diagnosis of canine influenza and the preventive therapy, and has a promising application prospect.

Description

technical field

[0001] The invention relates to a canine influenza virus monoclonal antibody hybridoma cell and belongs to the field of biotechnology. Background technique

[0002] Canine influenza (Canine influenza, CI) is a canine respiratory infectious disease caused by influenza A virus of the family Orthomyxoviridae. Influenza A virus has a wide infection spectrum and can infect humans, pigs, horses, poultry and other animals. Canine influenza virus (CIV) is mainly transmitted through the air, but it is also an important transmission route through the digestive tract or through contact with various items contaminated by the virus. In the past, it was generally believed that dogs were not susceptible to influenza A virus, but in the past ten years, it has been found that dogs can be infected by different subtypes of influenza virus. Affected dogs often have clinical symptoms such as sneezing, coughing, fever and runny nose, and the mortality rate is 1% to 5%. In 2004,...

Examples

Embodiment Construction

[0026] (1) Establishment of canine influenza virus monoclonal antibody hybridoma F112 strain

[0027] 1. Preparation of Canine Influenza Virus Immune Antigen

[0028] A / Canine / Nanjing / 11 / 2012(H3N2) (Wang Xiaoli, et al. Isolation and evolution analysis of a H3N2 subtype canine influenza virus. Chinese Journal of Animal Infectious Diseases, 2015,23(2):25-31) inoculated in 9-11 day-old SPF chicken embryos (purchased from Nanjing Tianbang Biotechnology Co., Ltd.), after 2-3 days, the hemagglutination titer is greater than 2 6 chicken embryo allantoic fluid. After the allantoic fluid was centrifuged at 5000r / min for 30min, the supernatant was taken, and then centrifuged at 8000r / min for 30min, and the supernatant was taken. After preliminary removal of impurities, ultracentrifuge at 30,000r / min for 3 hours, discard the supernatant, suspend the precipitate with an appropriate amount of PBS, prepare sucrose solutions of different concentrations, and use a pipette from high concentr...

technical field

[0001] The invention relates to a canine influenza virus monoclonal antibody hybridoma cell and belongs to the field of biotechnology. Background technique

[0002] Canine influenza (Canine influenza, CI) is a canine respiratory infectious disease caused by influenza A virus of the family Orthomyxoviridae. Influenza A virus has a wide infection spectrum and can infect humans, pigs, horses, poultry and other animals. Canine influenza virus (CIV) is mainly transmitted through the air, but it is also an important transmission route through the digestive tract or through contact with various items contaminated by the virus. In the past, it was generally believed that dogs were not susceptible to influenza A virus, but in the past ten years, it has been found that dogs can be infected by different subtypes of influenza virus. Affected dogs often have clinical symptoms such as sneezing, coughing, fever and runny nose, and the mortality rate is 1% to 5%. In 2004,...

Examples

Embodiment Construction

[0026] (1) Establishment of canine influenza virus monoclonal antibody hybridoma F112 strain

[0027] 1. Preparation of Canine Influenza Virus Immune Antigen

[0028] A / Canine / Nanjing / 11 / 2012(H3N2) (Wang Xiaoli, et al. Isolation and evolution analysis of a H3N2 subtype canine influenza virus. Chinese Journal of Animal Infectious Diseases, 2015,23(2):25-31) inoculated in 9-11 day-old SPF chicken embryos (purchased from Nanjing Tianbang Biotechnology Co., Ltd.), after 2-3 days, the hemagglutination titer is greater than 2 6 chicken embryo allantoic fluid. After the allantoic fluid was centrifuged at 5000r / min for 30min, the supernatant was taken, and then centrifuged at 8000r / min for 30min, and the supernatant was taken. After preliminary removal of impurities, ultracentrifuge at 30,000r / min for 3 hours, discard the supernatant, suspend the precipitate with an appropriate amount of PBS, prepare sucrose solutions of different concentrations, and use a pipette from high concentr...