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As a well-known powerful mucosal adjuvant peptides,Escherichia coli heat-labile enterotoxin(LT),especially the non-toxic or low toxic mutants(LTm),is capable of promoting strong mucosal immune responses to co-administered antigens in many types of vaccines[1-2].To prepare LTm efficiently,we have developed a novel method that expressing and purifying LTA,LTB subunits respectively,controlling two subunits proportion and simulating the environment in vivo to assembly them into hexamer structure.