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Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor
Center of Biological Defense, Těchonín, Czech Republic
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
Abstract: 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.
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MDPI and ACS Style
Pohanka, M.; Pavliš, O.; Skládal, P. Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor. Sensors 2007, 7, 341-353.
Pohanka M, Pavliš O, Skládal P. Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor. Sensors. 2007; 7(3):341-353.
Pohanka, Miroslav; Pavliš, Oto; Skládal, Petr. 2007. "Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor." Sensors 7, no. 3: 341-353.