Preparation of anti-morphine monoclonal antibodies with complete cross reactivity with heroin

2005 
OBJECTIVE: To prepare and characterize anti-morphine monoclonal antibodies (mAbs) with complete cross reactivity with heroin. METHODS: Morphine-6-succinyl was conjugated with Blue carrier (BC) in the presence of carbodi imide. BALB/c mice were immunized with the holoantigens (M-6-S-BC) and Freund adjuvant, and hybridoma was generated with mouse spleen cells and Sp2/0 myeloma cells. After 3 fusions and cloning, 4 hybridoma cell lines secreting monoclonal antibodies to morphine were obtained and the mABs purified from the mouse ascites followed by characterization with enzyme-linked immunosorbent assay (ELISA). RESULTS: Four hybridoma cell lines secreting anti-morphine mAbs were established and designated as G1, G3, H4 and B1, respectively. All the mAbs belonged to IgG1 isotype and could bind to heroin better than to morphine and codeine, which were identified by competitive inhibition ELISA. CONCLUSION: The anti-morphine mAbs can bind with the opioids, especially heroin, and can be useful for sensitive detection of opioids.
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