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Measuring proteins with greater speed and resolution while reducing sample size

A multi-angle light scattering (MALS) system, combined with chromatographic separation, directly measures the absolute molar mass, size and concentration of the eluate species. The measurement of these crucial properties in solution is essential in basic macromolecular characterization and all resea...

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Autores principales: Hsieh, Vincent H., Wyatt, Philip J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5577338/
https://www.ncbi.nlm.nih.gov/pubmed/28855530
http://dx.doi.org/10.1038/s41598-017-09051-1
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author Hsieh, Vincent H.
Wyatt, Philip J.
author_facet Hsieh, Vincent H.
Wyatt, Philip J.
author_sort Hsieh, Vincent H.
collection PubMed
description A multi-angle light scattering (MALS) system, combined with chromatographic separation, directly measures the absolute molar mass, size and concentration of the eluate species. The measurement of these crucial properties in solution is essential in basic macromolecular characterization and all research and production stages of bio-therapeutic products. We developed a new MALS methodology that has overcome the long-standing, stubborn barrier to microliter-scale peak volumes and achieved the highest resolution and signal-to-noise performance of any MALS measurement. The novel design simultaneously facilitates online dynamic light scattering (DLS) measurements. As National Institute of Standards and Technology (NIST) new protein standard reference material (SRM 8671) is becoming the benchmark molecule against which many biomolecular analytical techniques are assessed and evaluated, we present its measurement results as a demonstration of the unique capability of our system to swiftly resolve and measure sharp (20~25 µL full-width-half-maximum) chromatography peaks. Precise measurements of protein mass and size can be accomplished 10 times faster than before with improved resolution. In the meantime the sample amount required for such measurements is reduced commensurately. These abilities will have far-reaching impacts at every stage of the development and production of biologics and bio-therapeutic formulations.
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spelling pubmed-55773382017-09-06 Measuring proteins with greater speed and resolution while reducing sample size Hsieh, Vincent H. Wyatt, Philip J. Sci Rep Article A multi-angle light scattering (MALS) system, combined with chromatographic separation, directly measures the absolute molar mass, size and concentration of the eluate species. The measurement of these crucial properties in solution is essential in basic macromolecular characterization and all research and production stages of bio-therapeutic products. We developed a new MALS methodology that has overcome the long-standing, stubborn barrier to microliter-scale peak volumes and achieved the highest resolution and signal-to-noise performance of any MALS measurement. The novel design simultaneously facilitates online dynamic light scattering (DLS) measurements. As National Institute of Standards and Technology (NIST) new protein standard reference material (SRM 8671) is becoming the benchmark molecule against which many biomolecular analytical techniques are assessed and evaluated, we present its measurement results as a demonstration of the unique capability of our system to swiftly resolve and measure sharp (20~25 µL full-width-half-maximum) chromatography peaks. Precise measurements of protein mass and size can be accomplished 10 times faster than before with improved resolution. In the meantime the sample amount required for such measurements is reduced commensurately. These abilities will have far-reaching impacts at every stage of the development and production of biologics and bio-therapeutic formulations. Nature Publishing Group UK 2017-08-30 /pmc/articles/PMC5577338/ /pubmed/28855530 http://dx.doi.org/10.1038/s41598-017-09051-1 Text en © The Author(s) 2017 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/.
spellingShingle Article
Hsieh, Vincent H.
Wyatt, Philip J.
Measuring proteins with greater speed and resolution while reducing sample size
title Measuring proteins with greater speed and resolution while reducing sample size
title_full Measuring proteins with greater speed and resolution while reducing sample size
title_fullStr Measuring proteins with greater speed and resolution while reducing sample size
title_full_unstemmed Measuring proteins with greater speed and resolution while reducing sample size
title_short Measuring proteins with greater speed and resolution while reducing sample size
title_sort measuring proteins with greater speed and resolution while reducing sample size
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5577338/
https://www.ncbi.nlm.nih.gov/pubmed/28855530
http://dx.doi.org/10.1038/s41598-017-09051-1
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