Mb. Medina, HYGROMYCIN-B ANTIBODY-PRODUCTION AND CHARACTERIZATION BY A SURFACE-PLASMON RESONANCE BIOSENSOR, Journal of agricultural and food chemistry, 45(2), 1997, pp. 389-394
Sensitive and accurate methods are needed for the detection of hygromy
cin B antibiotic in fluids and tissues of farm animals. Sheep antisera
were produced from hygromycin B-keyhole limpet hemocyanin and were sc
reened with immunodiffusion, ELISA, and fluorescent latex assays. The
antisera were evaluated with the BIAcore, a surface plasmon resonance
biosensor, for their binding properties without using signal-generatin
g labels. Hygromycin B was immobilized on the sensor chip, and the cap
ture (binding) of the antibody resulted in a proportional increase in
mass. Evaluation of the association (k(a)) and dissociation rate (k(d)
) contants showed that one antibody had an affinity constant (k(a)/k(d
)) of 1.64E+10. The binding capacities and antisera specificity were d
etermined using a competitive binding of the added drug and hygromycin
sensor, detecting hygromycin B from 2.5 ng/mL to 5 mg/mL. Neomycin, g
entamicin, spectinomycin, dihydrostreptomycin, and streptomycin (1000
times above safe levels) had negligible binding with the antisera. The
BIAcore analysis was more rapid and accurate than the immunochemical
assays and allow rapid development of methods of hygromycin B analysis
in biological samples.