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Clinical Chemistry 35: 1026-1031, 1989;
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Clinical Chemistry, Vol 35, 1026-1031, Copyright © 1989 by American Association for Clinical Chemistry

Dissociation of immunocomplexes by ionic shock for the development of immunosensors: application to measurement of alpha 1-fetoprotein

JL Boitieux, MP Biron, G Desmet and D Thomas
Laboratoire de Technologie Enzymatique, Universite de Technologie, Compiegne, France.

We report here our experimental results on the reaction rate of immunological complexes and on potential repeated use of membranes as a result of dissociation of the complexes between alpha 1-fetoprotein and catalase-labeled antibodies by different buffers with various ionic strengths. The measurement consists of an immunological process and an enzymatic reaction. The protein membrane activated by glutaraldehyde for the immobilization of antibodies is fixed over an oxygen electrode. After incubation with antibody/alpha 1-fetoprotein and antibody/alpha 1- fetoprotein antibodies coupled to catalase, the reaction medium is introduced into a continuous-flow cell. Oxygen production by the catalase is measured on-line, with the electrode in contact with hydrogen peroxide. This response is correlated to the alpha 1- fetoprotein concentration of the sample. We show a typical calibration curve between 0.5 and 120 micrograms/L. Replicate (n = 20) equilibrium measurement with the same membrane gave a CV of 2.2%. The reversible immunochemical sensor has been tested for 30 measurements without significant loss of activity.




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Copyright © 1989 by the American Association for Clinical Chemistry.