DEV gB protein monoclonal antibody and blocking ELISA kit for detecting DEV antibody
A detection kit, monoclonal antibody technology, applied in the direction of antibodies, antiviral agents, immunoglobulins, etc., can solve the lack of blocking ELISA kits and other problems, achieve good commercial application prospects, high sensitivity, repeatability strong effect
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Embodiment 1
[0053] Example 1 Prokaryotic expression of DEV gB protein
[0054] 1. Materials and methods
[0055] 1.1 Virus strains and experimental animals
[0056] DEVCSC strain seed virus was preserved by our laboratory; SPF duck embryos were purchased from the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; clean grade New Zealand white rabbits were purchased from Liaoning Changsheng Biotechnology Co., Ltd.; DEV positive serum reference materials were purchased from China Veterinary Drug Supervision Institute; pET-32a vector plasmid is preserved by our laboratory.
[0057] 1.2 Test method
[0058] 1.2.1 Proliferation of DEV
[0059] Using conventional methods, duck embryo fibroblasts (DEFs) were prepared from 11-day-old SPF duck embryos, and the prepared primary cells were cultured in DMEM containing 5% FBS, and the primary cells were placed in 5% CO 2 In a 37°C incubator, subculture after 1-2 days, and grow to 80% of th...
Embodiment 2
[0129] Example 2 Preparation and selection of DEV and DEV gB protein monoclonal antibody
[0130] 1. Test material
[0131] 1.1 Virus strains and cells
[0132] H5, H7 and H9 subtype avian influenza virus (AIV) HI test antigens were purchased from Harbin Veken Biotechnology Development Company, Newcastle disease virus (NDV), egg drop syndrome virus (EDSV), duck Tembusu virus (DTMUV) Duck hepatitis virus type Ⅰ (DHVI Ⅰ) are all preserved by our laboratory. SP2 / 0 myeloma cells are maintained by the laboratory.
[0133] 1.2 Experimental animals and serum samples
[0134] Eleven-day-old SPF duck embryos were purchased from the Experimental Animal Center of Harbin Veterinary Research Institute. Six-week-old SPF female BALB / c mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. 6 positive serum samples were prepared from immunized SPF ducks in the laboratory and identified by neutralization test; 6 negative serum samples were collected from no...
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