Development of Rapid One-Step Immunochromatographic Assay
| Autor: | Seung Hwa Lee, Young-Sang Kim, Se-Hwan Paek, Joung-Hwan Cho |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Salmonella typhimurium
Analyte Time Factors Capillary action Gold Colloid Sensitivity and Specificity Antibodies General Biochemistry Genetics and Molecular Biology Epitope Antigen-Antibody Reactions chemistry.chemical_compound medicine Animals Antigens Molecular Biology Immunoassay Chromatography medicine.diagnostic_test Chemistry Goats Temperature Collodion Membrane Colloidal gold Reagent Thermodynamics Nitrocellulose |
| Zdroj: | Methods. 22:53-60 |
| ISSN: | 1046-2023 |
| DOI: | 10.1006/meth.2000.1036 |
| Popis: | An analytical system for a one-step immunoassay has been constructed using the concept of immunochromatography. The system employed two different antibodies that bound distinct epitopes of an analyte molecule: an antibody labeled with a signal generator (e.g., colloidal gold), which was placed in the dry state at a predetermined site on a glass-fiber membrane, and another antibody immobilized on the surface of a nitrocellulose membrane. Three membranes, one with the tracer, one with immobilized antibody, and a cellulose membrane as the absorbent of medium (in a sequence from the bottom), were attached to a plastic film and cut into strips. Aqueous medium containing analyte absorbed from the bottom end of the immunostrip dissolved the labeled antibody, and the antigen–antibody binding complex formed was transported into the next nitrocellulose membrane by the flow caused by capillary action. The complex subsequently reacted with the immobilized antibody, which generated a signal in proportion to the analyte concentration. The convective mass transfer of the immunoreactant to the binding partner allowed the assay to be performed with no handling of reagents. The reaction, however, was carried out under nonequilibrium conditions, which resulted in decreased sensitivity as compared with assays performed in an equilibrium mode (e.g., ELISA). To minimize such sacrifice, major factors that control system performance were identified and the system was then devised under optimal conditions. |
| Databáze: | OpenAIRE |
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