Anti-unfolded acetylcholinesterase monoclonal antibodies

The reduced and alkylated unfolded acetylcholinesterase(RA-AChE)protein was used as the antigen to immunize Balb/c mice by two injections for the basic immunization and three for the enhanced immunization.The splenocytes were fused with SP2/0-Agl4 mouse myeloma cells by PEG 1000.Five hybridoma cell lines secreting anti-RA-AChE monoclonal antibodies have been cloned.Antibody titers in the supernatants of the incubation media rose to 1:103 and in the ascites of the mice inoculated thehybridoma cells,1:105-6.The epitope analysis showed that the five monoclonal antibodies directed to three epitopes altogether on the acetykholinesterase molecule.They were all extensively immunoreactive to the RA-AChE and native AChE,however,did not significantly inhibit the enzyme activity.