An iBody-based lateral flow assay for semi-quantitative determination of His-tagged protein concentration

0509610 - ÚOCHB 2020 RIV NL eng J - Journal Article
Kryštůfek, Robin - Šácha, Pavel
An iBody-based lateral flow assay for semi-quantitative determination of His-tagged protein concentration.
Journal of Immunological Methods. Roč. 473, Oct (2019), č. článku 112640. ISSN 0022-1759. E-ISSN 1872-7905
R&D Projects: GA ČR(CZ) GA19-10280S; GA MPO(CZ) FV10490; GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : lateral flow assay * His-tag * antibody mimetics * protein quantitation
OECD category: Biochemical research methods
Impact factor: 1.901, year: 2019
Method of publishing: Limited access
https://www.sciencedirect.com/science/article/abs/pii/S0022175919302091?via%3Dihub

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.
Permanent Link: http://hdl.handle.net/11104/0300300