Preparation Of Monoclonal Antibodies Against CSFV And The Immunohistochemistry Identification

ABSTRACT The hybridoma cell lines,NF1,NF2,NF3,NF4 secreting anti-CSFV monoclonal antibodies were established by immunization of BALB/C mice with purified CSFV SM strain,using cell fusion technique.The number of chromosome are 96 to 115 pieces.The ELISA titers of NF2 cell supernatant and ascites were 1:80 and 1:10000.The monoclonal antibodies are specific to CSFV and can't bind to the normal samples such as pig's kidney and liver in indirect-ELISA and sandwich-ELISA.NF1,NF2,NF3,and NF4 all belong to class IgG,and subclass IgG1/λ,,IgG2a/κ,IgG2a/λ,IgG1/λrespectively.The results indicate that four hybridoma cell lines can infinitely secrete anti- CSFV monoclonal antibody that can be used as immunology reagent to develop sensitive,specific,fast diagnosis kit and reagent paper probe.A strain of McAb,named NF2 with isotype of IgG2a/κin mouse IgG subclass, show specific reaction to the CSFV infected PK-15 cell lines and the tissues of kidney and liver of pigs,but no response to that of uninfection,by indirect ELISA and immunohistochemistry identification.The ELISA titres of the McAb were 1:160 and 1:10000,in hydridoma cell supematant or mouse ascites respectively.The McAb had performed a bright immunostaining to CSFV in PK-15 cell lines and endotheliocytes of small vessels in the tissues.The result had confirmed that the McAb is a specific reagent for detection of the CSFV distribution in the host.