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Chromophore pre-mat...
Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion
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- Lundqvist, Magnus (author)
- KTH,Proteinteknologi
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- Thalén, Niklas (author)
- KTH,Proteinteknologi
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- Volk, Anna-Luisa (author)
- KTH,Proteinteknologi
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- Hansen, Henning Gram (author)
- Tech Univ Denmark, Novo Nordisk Fdn Ctr Biosustainabil, Lyngby, Denmark.
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- von Otter, Eric (author)
- Nanyang Technol Univ, Sch Biol Sci, Singapore 637551, Singapore.
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- Nygren, Per-Åke, 1961- (author)
- KTH,Proteinvetenskap
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- Uhlén, Mathias (author)
- KTH,Science for Life Laboratory, SciLifeLab,Proteinvetenskap,The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark
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- Rockberg, Johan (author)
- KTH,Proteinvetenskap
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(creator_code:org_t)
- 2019-01-22
- 2019
- English.
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In: Scientific Reports. - : NATURE PUBLISHING GROUP. - 2045-2322. ; 9
- Related links:
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https://www.nature.c...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- Complementation-dependent fluorescence is a powerful way to study co-localization or interactions between biomolecules. A split-GFP variant, involving the self-associating GFP 1-10 and GFP 11, has previously provided a convenient approach to measure recombinant protein titers in cell supernatants. A limitation of this approach is the slow chromophore formation after complementation. Here, we alleviate this lag in signal generation by allowing the GFP 1-10 chromophore to mature on a solid support containing GFP 11 before applying GFP 1-10 in analyses. The pre-maturated GFP 1-10 provided up to 150-fold faster signal generation compared to the non-maturated version. Moreover, pre-maturated GFP 1-10 significantly improved the ability of discriminating between Chinese hamster ovary (CHO) cell lines secreting GFP 11-tagged erythropoietin protein at varying rates. Its improved kinetics make the pre-maturated GFP 1-10 a suitable reporter molecule for cell biology research in general, especially for ranking individual cell lines based on secretion rates of recombinant proteins.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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