Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/103599
Title: Engineering protein thermostability using a generic activity-independent biophysical screen inside the cell
Authors: Asial, Ignacio
Cheng, Yue Xiang
Engman, Henrik
Dollhopf, Maria
Wu, Binghuang
Nordlund, Pär
Cornvik, Tobias
Keywords: DRNTU::Science::Biological sciences
Issue Date: 2013
Source: Asial, I., Cheng, Y. X., Engman, H., Dollhopf, M., Wu, B., Nordlund, P., & Cornvik, T. (2013). Engineering protein thermostability using a generic activity-independent biophysical screen inside the cell. Nature Communications, 4.
Series/Report no.: Nature communications
Abstract: Protein stability is often a limiting factor in the development of commercial proteins and biopharmaceuticals, as well as for biochemical and structural studies. Unfortunately, identifying stabilizing mutations is not trivial since most are neutral or deleterious. Here we describe a high-throughput colony-based stability screen, which is a direct and biophysical read-out of intrinsic protein stability in contrast to traditional indirect activity-based methods. By combining the method with a random mutagenesis procedure, we successfully identify thermostable variants from 10 diverse and challenging proteins, including several biotechnologically important proteins such as a single-chain antibody, a commercial enzyme and an FDA-approved protein drug. We also show that thermostabilization of a protein drug using our approach translates into dramatic improvements in long-term stability. As the method is generic and activity independent, it can easily be applied to a wide range of proteins.
URI: https://hdl.handle.net/10356/103599
http://hdl.handle.net/10220/19332
ISSN: 2041-1723
DOI: http://dx.doi.org/10.1038/ncomms3901
Rights: © 2013 Macmillan Publishers Limited.
metadata.item.grantfulltext: none
metadata.item.fulltext: No Fulltext
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