Abstract
On- and off-line heterogeneous non-competitive flow immunoassays for the determination of Interleukin-10 are described. The sample containing IL-10 is mixed, either on-line in a reaction coil or off-line in a test tube, with fluorescent labelled anti-IL-10 antibodies to form an antibody-antigen complex. The labelled unbound antibodies are trapped on an immobilized IL-10 column whereas the IL-10-antibody complexes are eluted and detected downstream by a fluorescence detector. The optimization of the systems was performed with respect to choice of affinity support, flow rate, carrier buffer additives, pH and antibody-antigen association. Both bio recognition assays were tested with a spiked cell medium and the IL-10 detection limits in this matrix was found to be 8 fmol using the off-line incubation mode and 40 fmol using the on-line incubation mode. The sample through-put was 26 and 40 samples per hour in the on-line and off-line incubation modes, respectively. IL-10 identification in the sample fractions was achieved using MALDI-TOF MS.
Original language | English |
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Pages (from-to) | 119-30 |
Journal | Journal of Immunological Methods |
Volume | 246 |
Issue number | 1-2 |
DOIs | |
Publication status | Published - 2000 Dec 1 |
Subject classification (UKÄ)
- Medicinal Chemistry
Free keywords
- Antigen-Antibody Reactions
- Chromatography, Affinity
- Cross Reactions
- Fluoroimmunoassay/methods
- Humans
- Interleukin-10/analysis
- Kinetics
- Recombinant Proteins/analysis
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Substrate Specificity