Optimization of mouse immunity in the preparation of Echinococcusgranulosus monoclonal antibody by the orthogonal design and central-composite design-response surface methodology

Abstract

Objective To optimize the immune strategy for the preparation of monoclonal antibody against Echinococusgranulosus. Methods The titer of the anti-serum measured by enzyme-linked immunosorbent assay (ELISA) , then the titer of the anti-serum and serum OD values have been used as a selection criterion to observe the effect of dosages of streptomycin, mouse reproductive algebra and immune injection method on antiserum titre by orthogonal experiments.Furthermore, box-behnken design-response surface methodology (BBD-RSM) method was used to investigate the effects of antigen dosage, mouse day age, intervals between first and second immunization on antiserum potency after immunization. Results The optimum option(0 IU of green streptocindose, subcutaneous injection, less than 5 of mouse reproductive generations) has been defined by the orthogonal design .While BD-RSM indicated that the best immune condition was 60 μg dose of antigen, 42 days old mouse and 15 days interval between the first and the second immunization. Conclusion The optimized immunization schemes of mouse in the preparation of Echinococcusgranulosus monoclonal antibody may be used the selection of immune mouse scheme. Key words: Echinoccocusgranucosus; Monoclnal antibody; Central-composite design-response surface methodology; orthogonal design