| Abstract
| - Sensitive and accurate methods are needed for the detection ofhygromycin B antibiotic in fluidsand tissues of farm animals. Sheep antisera were produced fromhygromycin B−keyhole limpethemocyanin and were screened with immunodiffusion, ELISA, andfluorescent latex assays. Theantisera were evaluated with the BIAcore, a surface plasmon resonancebiosensor, for their bindingproperties without using signal-generating labels. Hygromycin Bwas immobilized on the sensorchip, and the capture (binding) of the antibody resulted in aproportional increase in mass.Evaluation of the association (ka) anddissociation rate (kd) contants showed that oneantibody hadan affinity constant(ka/kd) of 1.64E+10.The binding capacities and antisera specificity weredetermined using a competitive binding of the added drug and hygromycinsensor, detectinghygromycin B from 2.5 ng/mL to 5 mg/mL. Neomycin, gentamicin,spectinomycin, dihydrostreptomycin, and streptomycin (1000 times above safe levels) had negligiblebinding with the antisera.The BIAcore analysis was more rapid and accurate than theimmunochemical assays and allowrapid development of methods of hygromycin B analysis in biologicalsamples. Keywords: Hygromycin; biosensor; anti-hygromycin; antibody production;surface plasmonresonance; BIAcore; ELISA; fluorescent latex assay
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