An iBody-based lateral flow assay for semi-quantitative determination of His-tagged protein concentration
| Autor: | Pavel Šácha, Robin Kryštůfek |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Immunology Quantitative proteomics Antibodies Epitope 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Western blot Limit of Detection medicine Immunology and Allergy Histidine Polyhistidine-tag Reagent Strips Detection limit Chromatography biology medicine.diagnostic_test Chemistry Proteins Protein engineering 030104 developmental biology Reagent biology.protein Antibody 030215 immunology |
| Zdroj: | Journal of Immunological Methods. 473:112640 |
| ISSN: | 0022-1759 |
| Popis: | The polyhistidine tag (His-tag) is one of the most commonly used epitope tags in protein engineering. While His-tagged proteins can be detected reliably using immunological methods such as ELISA and Western blot, these methods are costly and time-intensive, necessitating more facile solutions for preliminary qualitative determination and concentration estimation. To this end, we present a rapid test strip assay based on iBody antibody mimetics that target the His-tag. We compare this strategy to commercial antibody-based assays and discuss the advantages and caveats of lateral flow assay design. Our test strip detected a panel of His-tagged proteins with different tag attachment strategies with a visual detection limit of 1 μM and densitometric detection limit of 0.5 μM. Due to its chemical nature, the presented assay exhibits wide reagent compatibility in comparison to antibody-based assays. |
| Databáze: | OpenAIRE |
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