Establishment and identification of a mouse hybridoma cell line secreting monoclonal antibody against chaperonin GroEL in Escherichia coli cells

Objective To prepare the monoclonal antibody against chaperonin GroEL,a member of heat shock protein(hsp60)family,in Escherichia coli cells.Methods Mice were immunized with E.coli cell lysate.The hybridomas secreting monoclonal antibodies(mAb)against E.coli cell lysate were developed.The immunoprecipitation,SDS-PAGE,and mass spectrometry analysis were used for identification of mAb against chaperonin GroEL.Results Among 33 hybridomas secreting mAbs against E.coli cell proteins,mAb 1A5 could recognize a protein with 60 kDa in E.coli cell lysate,which was confirmed as chaperonin GroEL by mass spectrometry assays.Conclusion The monoclonal antibody 1A5 against chaperonin GroEL of E.coli is prepared successfully,which could provide a useful tool for study on the structure and function of chaperonin GroEL.