Anti-avian leukosis virus p27 protein monoclonal antibody, gold-colloidal strip containing same and application
A technology of avian leukosis virus and monoclonal antibody, which is applied in the fields of monoclonal antibody, gold standard test strip, and monoclonal antibody against p27 protein of avian leukosis virus, can solve the problem that there are no effective control measures for avian leukosis, and achieve easy promotion, High efficiency, clear and easy-to-discern results
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Embodiment 1
[0025] Example 1 Preparation of Anti-Avian Leukosis Virus p27 Protein Monoclonal Antibody and Analysis of Monoclonal Antibody Specificity
[0026] 1 Materials and methods
[0027] 1.1 Virus strains, cells and experimental animals
[0028] ALV-J (HB09JY03) was isolated and identified by Harbin Veterinary Research Institute. ALV-J (HPRS103, GenBank: Z46390), ALV-A (RAV-1, GenBank: M19113) and ALV-B (RAV-2, GenBank: M14902) were donated by Professor Venugopal Nair, Pirbright Institute, UK; DF-1, S / P20 cells were preserved by our laboratory; 6-week-old BALB / c female mice were purchased from the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences.
[0029] 1.2 Main reagents
[0030] Escherichia coli DH5α and BL21 competent, prokaryotic expression vector pET-30a were preserved by our laboratory; pMD-18T vector and IPTG were purchased from Bao Biological Engineering (Dalian) Co., Ltd.; PEG-4000, HPR-labeled goat anti-mouse ...
Embodiment 2
[0063] Embodiment 2 detects the preparation and properties of the gold standard test paper strip of avian leukosis virus
[0064] 1.1 Preparation of colloidal gold solution
[0065] Colloidal gold particles with a particle size of 25 nm were prepared by sodium citrate reduction method. Under stirring, 100mL of 0.01% chloroauric acid aqueous solution was heated to boiling, and 1.5mL of 1% sodium citrate aqueous solution was quickly added, and the color of the solution was observed to change from white to black and then to wine red. When the color of the solution turns completely wine red, continue to heat for 5 minutes, then stop heating, cool naturally and store in the dark at 2-8°C.
[0066] 1.2 Preparation of gold-labeled conjugates
[0067] Gold conjugate labeling: add 0.1MK dropwise to 10mL colloidal gold solution 2 CO 3 0.1 mL, stirred for 5 min, added 200 μL of mouse monoclonal antibody 3H11 (prepared in Example 1) against avian leukosis virus p27 antigen, mixed for ...
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