Analysis of allergic components in tissue protein of eriocheir sinensis with different extraction and proocessing methods / 中国免疫学杂志

ABSTRACT

Objective:To analyze protein and allergic components in Eriocheir sinensis tissue with different extraction and processing methods , provide an optimal antigen extraction protocol for detecting specific IgE of Eriocheir sinensis.Methods: The proteins were extracted from tissue protein of Eriocheir sinensis with PBS extraction method ,acetone extraction method ,lysate extraction method,respectively;Extract heating and without heating tissue protein of Eriocheir sinensis with lysate extraction method .The total protein components were analyzed by SDS-PAGE and two-dimensional electrophoresis.The allergen components were identified by Western blot.Results:The binding strength of protein components extracted by different methods and sIgE in the immunoblot assay dif -fered.Protein components extracted by PBS extraction method were mainly 94,85,76,66,18 kD,by acetone extraction method were mainly 94,85,76,36 kD,while by lysate extraction method were mainly 200,125,51,43,38 kD.Protein components extracted by PBS extraction method reacted with patient serum were mainly 76,66,53,43,38 and 18 kD,by acetone extraction method were mainly 76, 66,53,43,38 and 18 kD,while by lysate extraction method are mainly 200,105,94,76,43,38 and 18 kD.Heating and without heating proteins and allergic components were similar.Conclusion:Lysate extraction method is more suitable for determination of specific IgE antigen preparation purpose.The binding activity of sIgE and heating or without heating allergen components is similar .