Preparation and identification of gentamicin and its polyclonal antibody

To develop immunoassay for gentamicin, the artificial antigen and polyclonal antibody were prepared in this study. Identified through ultraviolet spectrum (UV) and SDS-PAG, the immunogen (GM-BSA) and coating antigen (gm-ova) of gentamicin were synthesized by the method of carbodiimide successfully. GM-BSA was used to immune New Zealand white rabbits, and the antiserum was obtained. In this study, we utilize a notive method to immune animal and obtained serum volume much more than conventional method's. One indirect competitive ELISA method has been developed to detect the titer and sensitivity of the antiserum. The results show that the titer of antiserum was continuously improving and the IC50 was nearly 3 ng/mL. On this basis, With an concentration range from 0.3 to 243 ng/mL, the IC50 is 2.954 ng/mL, and the IC10 is 0.056 ng/mL. The results of this study provide a basis for the breakthrough of serum volume, and also is a good start for the development of gentamicin ELISA kit.