A rapid enzyme immunoassay for measurement of HBK in blood

An enzyme immunoassay has been developed for the measurement of HBK (4-amino-2-hydroxybutylyldibekacin) a new semisynthetic aminoglycoside antibiotic. Antisera were raised in rabbits by immunization with HBK conjugated to bovine serum albumin (BSA). 3'-Eno-HBK conjugated to alkaline phosphatase (ALP) was used as an enzyme labeled antigen. The antibody-bound drug was separated from free using goat anti-rabbit IgG serum. The assay can be completed within one hr by co-incubating the first and the second antibody. The present immunoassay allows detection of 10 ng HBK per ml of serum, and is applicable for monitoring HBK level in blood. HBK concentrations in human sera were determined by the immunoassay during and after infusion and the levels were compared with those determined by high performance liquid chromatography (HPLC) and antimicrobial assay.