Method for producing all-pig-source monoclonal antibody with single B cell PCR technology
A monoclonal antibody and B cell technology, applied in the biological field, to achieve the effect of good gene diversity and high efficiency
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[0063] Example 1 Establishment of a method for isolating whole-pig-derived anti-PEDV monoclonal antibodies using a single B cell PCR monoclonal antibody technique
[0064] 1. Animal immunity
[0065] Select 3 healthy Landrace piglets about the age of 35 days, numbering is respectively #241, #242, #243, uses the PEDV CV777 strain of 2ml respectively (10 4 TCID 50 / ml) Houhai acupoint injection (first immunization); at intervals of 2 weeks, use 2 times the dose of the first immunization to inactivate the virus before injecting Houhai acupoint (second immunization); after 2 weeks, use the same dose as the second immunization to isolate it in our laboratory The virulent PEDV strain LNCT2 strain was boosted. Monitor ELISA antibody and neutralizing antibody titers. Necropsy, anticoagulated blood and mesenteric lymph nodes were collected.
[0066] 2. Isolation and screening of single B lymphocytes:
[0067] PEDV antigen-specific porcine memory lymphocytes were selected by double...
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