Monoclonal antibody hybridoma cell 3B5 strain capable of secreting H protein for resisting canine distemper virus
A technology of monoclonal antibody and canine distemper virus, applied in antiviral immunoglobulin, antiviral agent, antibody, etc. Excellent chemical performance, enhanced ability to neutralize canine distemper virus, and improved neutralizing activity
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[0027] (1) Establishment of monoclonal antibody hybridoma cell lines
[0028] 1. Preparation of CDV antigen
[0029]Inoculate the CDV Onderstepoort strain (product of Holland Intervet International Co., Ltd.) into Vero cells (purchased from ATCC cell bank), collect the infection supernatant after the cells appear completely lesioned, freeze and thaw three times, and centrifuge at 8000r / min for 3 minutes. Remove cell debris; collect supernatant and add PEG 6000 (product of Sigma Company) to a final concentration of 9%, then sodium chloride was added to a final concentration of 3%, stirred at room temperature until dissolved, and precipitated overnight at 4°C. The next day, centrifuge at 8000r / min for 1 hour, discard the supernatant, and resuspend with 5mL TNE solution (pH7.8). Add the resuspension to the 60%-40%-20% sucrose gradient solution, 60000r / min ultracentrifugation for 3 hours, collect the layered protein bands (between 60%-40%), draw the virus bands, negative The vi...
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