Preparation and identification of the monoclonal antibody against smooth Brucella LPS.

To preparation monoclonal antibody(MAb) against smooth Brucella LPS,the B.melitensis M5-90 were inactivated and used to immunize BALB/c mice,Hybridomas were obtained by standard cell fusion techniques.Two cell strains secreting MAb against S-LPS,named 4G6 and 16C5,were obtained by I-ELISA screening with Brucella vaccine strains M5-90,S-19 and Y.enterocolitica O∶9 as antigens.MAb 16C5 was highly specific and had no cross reaction with all Gram-negative bacteria tested,while MAb 4G6 had no cross reaction with all but Y.enterocolitica O∶9.MAb 4G6 belonged to IgM while MAb 16C5 belonged to IgG3,and both light chains belonged to κ.Ascites of MAb 16C5 was purified by protein G and successfully used in competitive ELISA to detect antibodies against Brucella in cattle serum.The highly sensitivee and specific MAb 16C5 would provide a potential agent for diagnosis of brucellosis .