A rapid method to produce anti-gentamicin antibody

The aminoglycoside antibiotic, gentamicin, was conjugated to erythrocytes or bovine serum albumin (BSA) by a simple procedure in which ECDI was employed as the coupling reagent. When rabbits were immunized by injecting gentamicin-goat erythrocyte conjugates, three kinds of antibody were produced: 1. anti-gentamicin antibody, 2. anti-ECDI antibody, 3. goat erythrocyte agglutinins. The interfering anti-ECDI antibody was easily neutralized by adding acidified ECDI solution to the immune serum. Goat agglutinins were avoided by employing rabbit erythrocytes as the carrier cell in the hemagglutination titration. Highly specific anti-gentamicin antiserum was produced in rabbits by first injecting an initial dose of gentamicin-BSA conjugate as an emulsion in incomplete Freund's adjuvant via the foot pad, followed by multiple intravenous injections of gentamicin-erythrocyte conjugates. The immunization took approximately 21 days. High titered anti-gentamicin antibody was also produced by foot pad inoculation of gentamicin-BSA conjugates; however, the time necessary to achieve comparable titers was considerably longer (55 days). The antibodies produced by both immunization procedures were mainly of the IgG class.