Kinetics of antigen binding to antibody microspots: strong limitation by mass transport to the surface
| Autor: | Igor Goychuk, Christoph Gauer, Konstantin V. Klenin, Jörg D. Hoheisel, Sven Rüffer, Wlad Kusnezow, Walter Sebald, Yana V. Syagailo |
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| Rok vydání: | 2005 |
| Předmět: |
Mass transport
Antibody microarray Kinetics Protein Array Analysis Biological Transport Active Antigen-Antibody Complex Biochemistry Models Biological Thyroglobulin Antigen-Antibody Reactions Microtiter plate Interferon-gamma Humans Antigens Molecular Biology Immunoassay Chromatography Binding Sites biology Chemistry Models Theoretical Antigen binding Receptor–ligand kinetics Orders of magnitude (mass) Hemocyanins Biophysics biology.protein Binding Sites Antibody Antibody Protein Binding |
| Zdroj: | Proteomics. 6(3) |
| ISSN: | 1615-9853 |
| Popis: | It is well documented that diffusion has generally a strong effect on the binding kinetics in the microtiter plate immunoassays. However, a systematic quantitative experimental evaluation of the microspot kinetics is still missing in the literature. Our work aims at filling this important gap of knowledge on the example of antigen binding to antibody microspots. A mathematical model was derived within the framework of two-compartment model and applied to the quantitative analysis of the experimental data obtained for typical antibody microspot assays. A strong mass-transport dependence of the antigen-antibody microspot kinetics was identified to be one of the main restrictions of this new technology. The binding reactions are slowed down in the microspot immunoassays by several orders of magnitude as compared with the corresponding well-stirred bulk reactions. The task to relax the mass-transport limitations should thus be one of the most important issues in designing the antibody microarrays. These limitations notwithstanding, the detection range of more than five orders of magnitude and the high sensitivity in the low femtomolar range were experimentally achieved in our study, demonstrating thus an enormous potential of this highly capable technology. |
| Databáze: | OpenAIRE |
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