Based on single B cell antibody technology preparing the all-dog canine distemper virus H protein monoclonal antibody
In order to study the function of the Hemagglutinin(H)protein of canine distemper virus(CDV)and to develop diagnostic reagents and therapeutic mAb against the H protein.In this study,the experimental dogs were immunized with CDV inactivated,and a single CDV H protein specific memory B cell from the peripheral blood was sorted by flow cytometry.Using Nestle PCR to amplify the variable region gene of the antibody.Transfect mammalian cells with heavy and light chain antibody plasmid,and the im-munogenicity of the antibody was identified by indirect ELISA method and IFA.The Nestle PCR results show that the efficiency of antibody heavy chain was 85.4%(82/96),and the amplification efficiency of light chain was 84.4%(81/96),finally 26 pairs of heavy and light chain gene were obtained by se-quencing analysis.There are 4 mAb with CDV H protein specificity were screened out by the indirect ELISA method.The IFA results show that the 4 mAbs caused specific fluorescence in Vero cells infected by CDV.In this study,the all-dog mAb of CDV H protein was prepared by single B cell antibody technology for the first time,which lays the foundation for the subsequent research on the function of CDV H protein,and provides a new idea for preparate other canine pathogens mAbs.
canine distemper virushemagglutinin proteinmAbsingle B cell antibody technique
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