A novel sensitive immunoassay method based on the Invader technique
| Autor: | Hiroko Kimura, Fukui Koichi, Kimikazu Hashino, Minjue Xie, Kazuko Matsumoto, Masanori Horie, Yuri Sakihama |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Streptavidin
Time Factors Biophysics Enzyme-Linked Immunosorbent Assay Biochemistry Horseradish peroxidase Sensitivity and Specificity Fluorescence chemistry.chemical_compound Europium medicine Fluorescence Resonance Energy Transfer Humans Fluorescein Molecular Biology Horseradish Peroxidase Fluorescent Dyes Detection limit Immunoassay Chromatography medicine.diagnostic_test biology Oligonucleotide Tumor Necrosis Factor-alpha Cell Biology Immunoassay method chemistry biology.protein |
| Zdroj: | Analytical biochemistry. 374(2) |
| ISSN: | 1096-0309 |
| Popis: | A novel and sensitive immunoassay method has been developed in which the conventional sandwich immunoassay and the highly sensitive DNA detection method, the Invader method, are combined. The signal amplification function of the latter method has been successfully used to enhance the sensitivity of the sandwich immunoassay. The new assay method may be called the Immuno-Invader assay. The assay format involves three important steps: (1) a target antigen is captured and flagged with a biotin-conjugated detection antibody by the sandwich method, (2) streptavidin and a biotin-conjugated oligonucleotide are added to form a complex with the detection antibody, and (3) the oligonucleotide in the complex is detected using the Invader method. The method was applied to the assay of human tumor necrosis factor-alpha (hTNF-alpha). Detection limits obtained were 0.1 pg/ml hTNF-alpha when a luminescent europium chelate was used with a time-resolved measurement mode, and 0.8 pg/ml when fluorescein was used with a normal prompt fluorescence measurement mode. On the other hand, the detection limit of a commercially available hTNF-alpha enzyme-linked immunosorbent assay that uses horseradish peroxidase was 3.5 pg/ml. These results demonstrate the feasibility and potential of the new assay method for highly sensitive immunoassay. |
| Databáze: | OpenAIRE |
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