Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 473, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jim.2019.112640
Keywords
Lateral flow assay; His-tag; Antibody mimetics; Protein quantitation
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Funding
- Czech Science Foundation [19-10280S]
- Ministry of Industry and Trade of the Czech Republic [FV10490]
- European Regional Development Fund
- OP RDE [CZ.02.1.01/0.0/0.0/16_019/0000729]
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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 WA strip detected a panel of His-tagged proteins with different tag attachment strategies with a visual detection limit of 1 mu M and densitometric detection limit of 0.5 mu M. Due to its chemical nature, the presented assay exhibits wide reagent compatibility in comparison to antibody-based assays.
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