| Objective: To study the enzyme activity variance of monoclonal antibody on anti-acetycholinestrerases antibody in cultivation superior schedul and Ascites and the clinical significance as a guideline of threrapy organophosphorus.Methods: AchE have a good quality of antigen as antigen to immune mice, preparation monoclonal antibody on AchE; AchE-Sepharose-4B and SPA affinity column are applied for the purification of AchE monoclonal antibody (3F3), Enzyme cut 3F3 with Papain, getting Fab fragment; indirect method of ELISA determine the Fab fragment; Immune mice. Preparation anti-3F3 Fab monoclonal antibody with hybridoma; use ELISA determine activity on antibody enzyme of acetycholinesterases.The mean comparision use T detection.Results: Preparation anti-Fab idiotype antibody and determine antibody have acetycholinestrerases activity. 1. AchE 3F3 Fab have good activity of combining antigen AchE and immuntigen 2.antibody enzyme of acetycholinesterases have acetycholinestrerases activity. 3.the significant difference in monoclonal antibody 4B9 and 2B1 of different dilution (P<0.05,P<0.01).Conclusion: 1.acetycholinesterases have good antigen qulity ,use this produce monoclonal antibody. 2.cut acetycholinesterases with Pepsin,getting Fab fragment as antigen to produce antigenic determinant antibody. 3. Antibody enzyme of acetycholinesterases in Ascites and cultivation superior schedul have activity of AchE, variant dilution cause different activity. Antibody enzyme of acetycholinesterases havehigher peculiar, selection and simper manipulation than traditional enzyme, observing activity provide a new reliable way for clinical therapy of organophosphorus.Postgraduate: Weiwei Zhan (Immunology)Directed by: Prof. Meihua Gao... |
