Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion

Magnus Lundqvist, Niklas Thalén, Anna Luisa Volk, Henning Gram Hansen, Eric von Otter, Per-Åke Nygren, Mathias Uhlén, Johan Rockberg*

*Corresponding author for this work

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Abstract

Complementation-dependent fuorescence 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 signifcantly 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.
Original languageEnglish
Article number310
JournalScientific Reports
Volume9
Issue number1
ISSN2045-2322
DOIs
Publication statusPublished - 2019

Cite this

Lundqvist, Magnus ; Thalén, Niklas ; Volk, Anna Luisa ; Hansen, Henning Gram ; von Otter, Eric ; Nygren, Per-Åke ; Uhlén, Mathias ; Rockberg, Johan. / Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
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title = "Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion",
abstract = "Complementation-dependent fuorescence 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 signifcantly 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.",
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year = "2019",
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language = "English",
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Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion. / Lundqvist, Magnus; Thalén, Niklas; Volk, Anna Luisa; Hansen, Henning Gram; von Otter, Eric; Nygren, Per-Åke; Uhlén, Mathias; Rockberg, Johan.

In: Scientific Reports, Vol. 9, No. 1, 310, 2019.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion

AU - Lundqvist, Magnus

AU - Thalén, Niklas

AU - Volk, Anna Luisa

AU - Hansen, Henning Gram

AU - von Otter, Eric

AU - Nygren, Per-Åke

AU - Uhlén, Mathias

AU - Rockberg, Johan

PY - 2019

Y1 - 2019

N2 - Complementation-dependent fuorescence 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 signifcantly 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.

AB - Complementation-dependent fuorescence 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 signifcantly 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.

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