Development of 48-condition buffer screen for protein stability assessment

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F21%3A00130284" target="_blank" >RIV/00216224:14740/21:00130284 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Development of 48-condition buffer screen for protein stability assessment

Popis výsledku v původním jazyce

The determination of a suitable buffer environment for a protein of interest is not an easy task. The choice of the buffer is important both for basic research and for scaling up for use in biotechnologies. However, the requirements of advanced techniques, the demands on the biological material, and the researcher time needed for buffer optimization, as well as personal inflexibility, lead frequently to the use of sub-optimal buffers. Here, we demonstrate the design of a 48-condition buffer screen that can be used to determine an appropriate environment for downstream studies [1]. By the combination of several techniques (differential scanning fluorimetry, dynamic light scattering, and bio-layer interferometry), we are able to assess the protein stability, homogeneity, and binding activity across the screen with as little as 100 μg of protein in 1 day. The application of this screen helps to avoid unsuitable conditions, explain problems observed upon protein analysis, and choose the most suitable buffers for further research and applications. The screen can be routinely used as a primary screen for buffer optimization in laboratories and facilities.

Název v anglickém jazyce

Development of 48-condition buffer screen for protein stability assessment

Popis výsledku anglicky

The determination of a suitable buffer environment for a protein of interest is not an easy task. The choice of the buffer is important both for basic research and for scaling up for use in biotechnologies. However, the requirements of advanced techniques, the demands on the biological material, and the researcher time needed for buffer optimization, as well as personal inflexibility, lead frequently to the use of sub-optimal buffers. Here, we demonstrate the design of a 48-condition buffer screen that can be used to determine an appropriate environment for downstream studies [1]. By the combination of several techniques (differential scanning fluorimetry, dynamic light scattering, and bio-layer interferometry), we are able to assess the protein stability, homogeneity, and binding activity across the screen with as little as 100 μg of protein in 1 day. The application of this screen helps to avoid unsuitable conditions, explain problems observed upon protein analysis, and choose the most suitable buffers for further research and applications. The screen can be routinely used as a primary screen for buffer optimization in laboratories and facilities.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10610 - Biophysics

Projekt

<a href="/cs/project/LM2018127" target="_blank" >LM2018127: Česká infrastruktura pro integrativní strukturní biologii</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů