Preparation and primary application of 14-3-3 specific polyclonal antibody / 中华神经科杂志

ABSTRACT

Objective Preparation of specific antibody of neuron protein 14 3 3 and application for the diagnosis of prion diseases. Methods The rabbits were immunized with prokaryotic expressed GST 14 3 3 fusion protein and the collected antiserum was evaluated by ELISA and Western blot assays. Results ELISA revealed that the reactive titer of the prepared antibody against purified 14 3 3 protein was up to 14 096 000; Western blot assays confirmed that both prokaryotic expressed,and the natively presenting 14 3 3 proteins in the brain extracts were recognized by the prepared antibody. Two cases of clinically diagnosed Creutzfeldt Jakob disease (CJD)with typical EEG changes showed the positive 14 3 3 protein in cerebral spinal fluid (CSF). Four patients with dementia and narcoma but without typical EEG change were 14 3 3 negative in CSF,three of them being even further tested by brain biopsies without typical histological change of CJD and presentation of protease resistant PrP Sc protein. 35 samples CFS collected from patients without dementia were 14 3 3 negative in CSF. All the results were the same as that of standard 14 3 3 polyclonal antibody.Conclusion Using purified prokaryotic expressed GST 14 3 3 fusion protein as antigen,the specific antibody was elicited in the immunized animals. The prepared antibody can be used in identification of 14 3 3 protein in CSF for the diagnosis of CJD.