Clinical Chemistry
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Clinical Chemistry 42: 1702-1708, 1996;
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Clinical Chemistry, Vol 42, 1702-1708, Copyright © 1996 by American Association for Clinical Chemistry

Analyte and label binding assay read by flow cytometry

JO Utgaard, J Frengen, T Stigbrand, A Ullen, R Schmid and T Lindmo
Department of Physics, Norwegian University of Science and Technology, Trondheim.

A new immunometric two-site sandwich assay is introduced, in which a label-scavenging binding partner is added to the sample in addition to the analyte-binding partner. The scavenger binding partner binds excess label antibody, giving a signal proportional to the amount of excess label antibody in the sample solution. A set of two calibration curves is obtained from the two binding partners simultaneously, and a combination of the two signals gives an unambiguous determination of the analyte concentration, even for high analyte concentrations where the hook effect may occur. Two-particle immunofluorometric assays developed for placental alkaline phosphatase and human chorionic gonadotropin on the basis of this principle and yielding signals measured by flow cytometry gave rapid results (2 h) and had working ranges in excess of 5 and 6 orders of magnitude for the respective analytes.


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C. Camilla, L. Mely, A. Magnan, B. Casano, S. Prato, S. Debono, F. Montero, J.-P. Defoort, M. Martin, and V. Fert
Flow Cytometric Microsphere-Based Immunoassay: Analysis of Secreted Cytokines in Whole-Blood Samples from Asthmatics
Clin. Vaccine Immunol., July 1, 2001; 8(4): 776 - 784.
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