Publication

A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables

Wang, Y., van Oosterwijk, N., Ali, A. M., Adawy, A., Anindya, A. L., Dömling, A. S. S. & Groves, M. R. 24-Aug-2017 In : Scientific Reports. 7, 1, 10 p., 9355

Research output: Scientific - peer-reviewArticle

APA

Wang, Y., van Oosterwijk, N., Ali, A. M., Adawy, A., Anindya, A. L., Dömling, A. S. S., & Groves, M. R. (2017). A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables. Scientific Reports, 7(1), [9355]. DOI: 10.1038/s41598-017-09687-z

Author

Wang, Yuanze; van Oosterwijk, Niels; Ali, Ameena M; Adawy, Alaa; Anindya, Atsarina L; Dömling, Alexander S S; Groves, Matthew R / A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables.

In: Scientific Reports, Vol. 7, No. 1, 9355, 24.08.2017.

Research output: Scientific - peer-reviewArticle

Harvard

Wang, Y, van Oosterwijk, N, Ali, AM, Adawy, A, Anindya, AL, Dömling, ASS & Groves, MR 2017, 'A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables' Scientific Reports, vol 7, no. 1, 9355. DOI: 10.1038/s41598-017-09687-z

Standard

A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables. / Wang, Yuanze; van Oosterwijk, Niels; Ali, Ameena M; Adawy, Alaa; Anindya, Atsarina L; Dömling, Alexander S S; Groves, Matthew R.

In: Scientific Reports, Vol. 7, No. 1, 9355, 24.08.2017.

Research output: Scientific - peer-reviewArticle

Vancouver

Wang Y, van Oosterwijk N, Ali AM, Adawy A, Anindya AL, Dömling ASS et al. A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables. Scientific Reports. 2017 Aug 24;7(1). 9355. Available from, DOI: 10.1038/s41598-017-09687-z


BibTeX

@article{9fb2b8ee66ea47f9ade8e1507e294e84,
title = "A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables",
abstract = "Refolding of proteins derived from inclusion bodies is very promising as it can provide a reliable source of target proteins of high purity. However, inclusion body-based protein production is often limited by the lack of techniques for the detection of correctly refolded protein. Thus, the selection of the refolding conditions is mostly achieved using trial and error approaches and is thus a time-consuming process. In this study, we use the latest developments in the differential scanning fluorimetry guided refolding approach as an analytical method to detect correctly refolded protein. We describe a systematic buffer screen that contains a 96-well primary pH-refolding screen in conjunction with a secondary additive screen. Our research demonstrates that this approach could be applied for determining refolding conditions for several proteins. In addition, it revealed which {"}helper{"} molecules, such as arginine and additives are essential. Four different proteins: HA-RBD, MDM2, IL-17A and PD-L1 were used to validate our refolding approach. Our systematic protocol evaluates the impact of the {"}helper{"} molecules, the pH, buffer system and time on the protein refolding process in a high-throughput fashion. Finally, we demonstrate that refolding time and a secondary thermal shift assay buffer screen are critical factors for improving refolding efficiency.",
keywords = "Journal Article",
author = "Yuanze Wang and {van Oosterwijk}, Niels and Ali, {Ameena M} and Alaa Adawy and Anindya, {Atsarina L} and Dömling, {Alexander S S} and Groves, {Matthew R}",
year = "2017",
month = "8",
doi = "10.1038/s41598-017-09687-z",
volume = "7",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",

}

RIS

TY - JOUR

T1 - A Systematic Protein Refolding Screen Method using the DGR Approach Reveals that Time and Secondary TSA are Essential Variables

AU - Wang,Yuanze

AU - van Oosterwijk,Niels

AU - Ali,Ameena M

AU - Adawy,Alaa

AU - Anindya,Atsarina L

AU - Dömling,Alexander S S

AU - Groves,Matthew R

PY - 2017/8/24

Y1 - 2017/8/24

N2 - Refolding of proteins derived from inclusion bodies is very promising as it can provide a reliable source of target proteins of high purity. However, inclusion body-based protein production is often limited by the lack of techniques for the detection of correctly refolded protein. Thus, the selection of the refolding conditions is mostly achieved using trial and error approaches and is thus a time-consuming process. In this study, we use the latest developments in the differential scanning fluorimetry guided refolding approach as an analytical method to detect correctly refolded protein. We describe a systematic buffer screen that contains a 96-well primary pH-refolding screen in conjunction with a secondary additive screen. Our research demonstrates that this approach could be applied for determining refolding conditions for several proteins. In addition, it revealed which "helper" molecules, such as arginine and additives are essential. Four different proteins: HA-RBD, MDM2, IL-17A and PD-L1 were used to validate our refolding approach. Our systematic protocol evaluates the impact of the "helper" molecules, the pH, buffer system and time on the protein refolding process in a high-throughput fashion. Finally, we demonstrate that refolding time and a secondary thermal shift assay buffer screen are critical factors for improving refolding efficiency.

AB - Refolding of proteins derived from inclusion bodies is very promising as it can provide a reliable source of target proteins of high purity. However, inclusion body-based protein production is often limited by the lack of techniques for the detection of correctly refolded protein. Thus, the selection of the refolding conditions is mostly achieved using trial and error approaches and is thus a time-consuming process. In this study, we use the latest developments in the differential scanning fluorimetry guided refolding approach as an analytical method to detect correctly refolded protein. We describe a systematic buffer screen that contains a 96-well primary pH-refolding screen in conjunction with a secondary additive screen. Our research demonstrates that this approach could be applied for determining refolding conditions for several proteins. In addition, it revealed which "helper" molecules, such as arginine and additives are essential. Four different proteins: HA-RBD, MDM2, IL-17A and PD-L1 were used to validate our refolding approach. Our systematic protocol evaluates the impact of the "helper" molecules, the pH, buffer system and time on the protein refolding process in a high-throughput fashion. Finally, we demonstrate that refolding time and a secondary thermal shift assay buffer screen are critical factors for improving refolding efficiency.

KW - Journal Article

U2 - 10.1038/s41598-017-09687-z

DO - 10.1038/s41598-017-09687-z

M3 - Article

VL - 7

JO - Scientific Reports

T2 - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 9355

ER -

ID: 47466356