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Gradient-free determination of isoelectric points of proteins on chip


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Publication Date: 2017-09-14

Journal Title: Physical Chemistry Chemical Physics

Publisher: Royal Society of Chemistry

Volume: 19

Pages: 23060-23067

Language: English

Type: Article

This Version: AM

Metadata: Show full item record

Citation: Łapińska, U., Saar, K. L., Yates, E., Herling, T., Müller, T., Challa, P. K., Dobson, C. M., & et al. (2017). Gradient-free determination of isoelectric points of proteins on chip. Physical Chemistry Chemical Physics, 19 23060-23067. https://doi.org/10.1039/c7cp01503h

Abstract: The isoelectric point (pI) of a protein is a key characteristic that influences its overall electrostatic behaviour. The majority of conventional methods for the determination of the isoelectric point of a molecule rely on the use of spatial gradients in pH, although significant practical challenges are associated with such techniques, notably the difficulty in generating a stable and well controlled pH gradient. Here, we introduce a gradient-free approach, exploiting a microfluidic platform which allows us to perform rapid pH change on chip and probe the electrophoretic mobility of species in a controlled field. In particular, in this approach, the pH of the electrolyte solution is modulated in time rather than in space, as in the case for conventional determinations of the isoelectric point. To demonstrate the general approachability of this platform, we have measured the isoelectric points of representative set of seven proteins, bovine serum albumin, $\beta$-lactoglobulin, ribonuclease A, ovalbumin, human transferrin, ubiquitin and myoglobin in microlitre sample volumes. The ability to conduct measurements in free solution thus provides the basis for the rapid determination of isoelectric points of proteins under a wide variety of solution conditions and in small volumes.

Sponsorship: We acknowledge financial support from the Biotechnology and Biological Sciences Research Council, the Newman Foundation and the Elan Pharmaceuticals. The research leading to these results has also received funding from the European Research Council under the European Union’s Seventh Framework Programme (FP7/2007- 2013) through the ERC grant PhysProt (agreement n1 337969).

Embargo Lift Date: 2100-01-01

Identifiers:

External DOI: https://doi.org/10.1039/c7cp01503h

This record's URL: https://www.repository.cam.ac.uk/handle/1810/265153



Rights: Attribution 4.0 International

Licence URL: http://creativecommons.org/licenses/by/4.0/





Autor: Łapińska, USaar, Kadi LiisYates, EVHerling, TWMüller, TChalla, Pavan KumarDobson, Christopher Martin Knowles, Tuomas PerttiSho

Fuente: https://www.repository.cam.ac.uk/handle/1810/265153



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