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ANTIGEN USED FOR IMMUNIZATIONWe used inactivated whole SARS CoV coronavirus as anti gen courtesy of Dr Anton Andonov National Microbiology Laboratory Winnipeg Canada METHOD OF IMUNIZATION For immunizations 5 6 week old female BALB C mice wereinjected subcutaneously with 50 mg of inactivated SARS CoVwith an equal part of Complete Freund s Adjuvant CFA H37Ra Difco on day 1 On day 30 the mice received 50 mg of purified SARS CoV in Incomplete Freund s Adjuvant IFA On days 48 and 63 mice received 5 mg in IFA The mice re ceived a final booster injection with 5 mg of purified SARSvirus 3 days prior to fusion Mice were euthanised by anaes thesia overdose and exsanguinated by cardiac puncture Thespleens were excised under aseptic conditions PARENTAL CELL LINE USED FOR FUSIONP3X63 Ag8 653 SELECTION AND CLONING OF HYBRIDOMASImmunization of mice removal of spleens preparation of spleen and myeloma cells fusion and screening for monoclonalantibody to SARS were performed according to NCFAD stan dard operating procedures under ISO17025 Hybridomas werecloned out in semisolid medium Supernatants were screened via ELISA using purified virus as antigen Isotyping was per formed using a commercial dipstick test Roche according tothe manufacturer s instructions SPECIFICITYNucleoprotein ISOTYPEIgG2b k SOURCEMurine FORMAT PURITYSupernatants IMMUNOGENWhole inactivated TOR 3 SARS CoV ANTIGEN REACTIVITYEIA Western APLICATIONSEIA Western cELISA METHOD OF IMUNIZATIONs q STORAGEFrozen 220 HYBRIDOMA AND HYBRIDOMICSVolume 23 Number 2 2004 Mary Ann Liebert Inc F26G15 Murine Monoclonal Antibody to the SARSCoronavirus Nucleoprotein SARS CoV 139 Monoclonal Antibodies MEDIABD Cell Quantum Yield AVAILABILITYSupernatant Yes3No ADRESS CORESPONDENCE TO Dr Jody D Berry Xin Yuan Brigitte Nicolas Fran Ranada and Frank Plummer The Monoclonal Antibody SectionNational Centre for Foreign Animal Disease CFIANational Microbiology Laboratory Health CanadaReagent Development Unit 1015 Arlington St Winnipeg MB CanadaTelephone 204 789 6063Fax 204 789 2038 E mail berryjd inspection gc ca MONOCLONAL ANTIBODIES140