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Sensors 2007, 7(3), 341-353; doi:10.3390/s7030341

Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor

1, 1 and 2,*
1 Center of Biological Defense, Těchonín, Czech Republic 2 Institute of Biochemistry, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic
* Author to whom correspondence should be addressed.
Received: 22 February 2007 / Accepted: 15 March 2007 / Published: 20 March 2007
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Monoclonal antibodies with specificity against the Francisella tularensis outerlipopolysaccharide (LPS) membrane were prepared and characterized using the piezoelectricimmunosensor with immobilized LPS antigen from F. tularensis. Signals obtained by theimmunosensor were compared with ELISA and similar sensitivity was noticed. Signal ofnegative controls obtained using the biosensor was below 0.5% of the signal obtained for theselected specific antibody clone 4H3B9D3. Testing of cross reactivity based on the sensorswith immobilized LPS from Escherichia coli and Bacillus subtilis confirmed selectivity ofthis antibody. Furthermore, the 4H3B9D3 antibody was successfully isotypized as IgM usingthe piezoelectric sensors with secondary antibodies. Kinetics parameters of antibody wereevaluated in the flow-through arrangement. The kinetic rate constants for the antibody4H3B9D3 were ka = (2.31 ± 0.20)·105 l mol-1s-1 (association) and kd = (0.0010 ±0.00062) s-1(dissociation) indicating very good affinity to the LPS antigen.
Keywords: piezoelectric resonator; immunosensor; Francisella tularensis; kinetic characterization. piezoelectric resonator; immunosensor; Francisella tularensis; kinetic characterization.
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Pohanka, M.; Pavliš, O.; Skládal, P. Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor. Sensors 2007, 7, 341-353.

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