Colloidal carbon particles as a new label for rapid immunochemical test methods: quantitative computer image analysis of results

A. van Amerongen, J.H. Wichers, L.B.J.M. Berendsen, A.J.M. Timmermans, G.D. Keizer, A.W.J. van Doorn, A. Bantjes, W.M.J. van Gelder

Research output: Contribution to journalArticleAcademic

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Abstract

Colloidal carbon particles can serve as label in sol particle immunoassays. The universal applicability of these particles in qualitative and (semi)quantitative immunoassays has been demonstrated. Sol particle and/or dipstick immunoassays, not yet optimized in terms of sensitivity, are discussed. The colloidal label has been used successfully in a mouse immunoglobulin isotyping kit. Human serum albumin spotted onto nitrocellulose in a concentration range of 7.8 to 1000 ng could be detected using anti-albumin antibody adsorbed onto colloidal carbon particles. It was also possible to perform a competitive assay with this conjugate for a concentration range of free human serum albumin varying from 0.25 to 6.75 ¿g. The Kunitz-type trypsin inhibitor from soybean was determined by a colloidal carbon based immunoassay in a range of 2.5 to 160 ng. In this assay, free and colloidal carbon-bound inhibitor competed for binding specific antibodies spotted onto a nitrocellulose membrane. An image- and data-processing procedure has been developed that enables a rapid and simple quantification of colloidal carbon sol particle immunoassays. The average grey level of a spot is taken as a measure for quantitative purposes. This so-called Sol-particle Image Processed ImmunoAssay (SIPIA) procedure is equally well applicable to assays using other colloidal particles.
Original languageEnglish
Pages (from-to)185-195
JournalJournal of biotechnology
Volume30
Issue number2
DOIs
Publication statusPublished - 1993

Keywords

  • IR-57415

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