Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/85425
Title: Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion
Authors: Lundqvist, Magnus
Thalén, Niklas
Volk, Anna-Luisa
Hansen, Henning Gram
von Otter, Eric
Nygren, Per-Åke
Uhlen, Mathias
Rockberg, Johan
Keywords: DRNTU::Science::Biological sciences
Chromophore
Protein Secretion
Issue Date: 2019
Source: Lundqvist, M., Thalén, N., Volk, A.-L., Hansen, H. G., von Otter, E., Nygren, P.-Å., . . . Rockberg, J. (2019). Chromophore pre-maturation for improved speed and sensitivity of split-GFP monitoring of protein secretion. Scientific Reports, 9, 310-. doi:10.1038/s41598-018-36559-x
Series/Report no.: Scientific Reports
Abstract: 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.
URI: https://hdl.handle.net/10356/85425
http://hdl.handle.net/10220/48210
DOI: 10.1038/s41598-018-36559-x
Rights: © 2019 The Author(s) (Nature Publishing Group). Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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