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?:abstract
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To explore the PED treatment preparation that can directly be used in piglets, here we inserted the PEDV neutralizingantibody PC10-IgG gene sequence into the human adenovirus type V (Ad5) backbone plasmid to construct the recombinantadenovirus Ad5-PC10-IgG and analyzed its anti-PEDV effect The supernatant of HEK293 cells infected with Ad5-PC10-IgG wasdetected by ELISA, and purified IgG from the supernatant was detected by SDS-PAGE and western blot The results showed that HEK293 cells infected by Ad5-PC10-IgG were able to secrete and express PC10-IgG and the antibody titer was relatively high Thesecreted PC10-IgG was also used to test the binding activity (by IFA) and neutralize activity (by PCR and IFA) against PEDV, andthe results showed that PC10-IgG expressed by Ad5-PC10-IgG in the supernatant could significantly inhibit PEDV infection in VeroE6 cells and had good binding activity In order to verify whether Ad5-PC10-IgG can infect the pig\'s small intestine, Ad5-PC10-IgGwere used to infect porcine intestinal enteroids, and the supernatant after infection was detected by ELISA as well as the expressionof PC10-IgG by porcine intestinal enteroids were detected by IFA The results showed that Ad5-PC10-IgG was able to infect porcineintestinal enteroids and the PC10-IgG was secreted by viral infected cells In addition, mice were inoculated by Ad5-PC10-IgG, andthe serum and anal swabs were collected at different time points and detected by ELISA After that, the binding activity andneutralizing activity of PC10-IgG in the serum against PEDV were detected through the virus binding experiment and the virusinhibition experiment The results showed that on the 1, 3, 5 day after inoculation, the serum of the inoculated mice contained higherconcentrations of PC10-IgG instead of anal swabs, and the PC10-IgG in the serum had good binding activity and neutralizingactivity to PEDV The above results indicate that Ad5-PC10-IgG can secrete and express the active neutralizing antibody PC10-IgGin vivo and in vitro Therefore, this study lays the foundation for the further development of PEDV therapeutic antibody againstPEDV infection in the future
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