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Since 1994, the use of chloramphenicol(CAP) is totally banned
within the European Union. As a consequence, levels for the
detection of CAP have to be changed. Piezoimmunosensor that
consists of selectivity and specificity of antigen antibody
interaction and sensitivity of mass change proportional to the
frequency shift was considered potential for CAP determination.
In this study, a water soluble carbodiimide was used to acylate
CAP succinate with bovine serum albumin. The conjugate retained
the bacteriostatic effect of CAP and fully functionized as an
immunogen. By fusing antibodies secreting cells with FO cells,
the hybridomas were developed. Five monoclones were colned with
limiting dilution method. All of them were classified as IgG1
containing light chain. Purified antibody was obtained by
Protein A affinity column chromatography. The measurement of
piezoelectric sensor was set up with a 10 MHz quartz crystal
oscillated by a SN 7400-TTL-IC and a universal counter connected
with personal computer through GBIP interface. The frequency
shifts can be monitored on PC and the sensitivity of
piezoimmunosensor is 1.7 Hz/ng by both-sides-open crystal bared
in air. Pretreated with Fc fragment specific anti-IgG, the gold
surfaces of quartz crystal can be immobilized with anti-CAP
oriently. The optimal condition for anti-IgG modification of
gold electrodes was investigated. On the basis of a competitive
immunoreaction with a protein-labelled CAP, the
piezoimmunosensor was developed with adequate sensitivity. The
decrease in frequency is inversely related to the CAP
concentration in the sample.
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