Development of 48-condition buffer screen for protein stability
assessment

J 2021

Development of 48-condition buffer screen for protein stability assessment

HOUSER, Josef, Jana KOSOUROVÁ, Monika KUBÍČKOVÁ and Michaela WIMMEROVÁ

Basic information

Original name

Development of 48-condition buffer screen for protein stability assessment

Authors

HOUSER, Josef (203 Czech Republic, guarantor, belonging to the institution), Jana KOSOUROVÁ (203 Czech Republic, belonging to the institution), Monika KUBÍČKOVÁ (203 Czech Republic, belonging to the institution) and Michaela WIMMEROVÁ (203 Czech Republic, belonging to the institution)

Edition

European Biophysics Journal With Biophysics Letters, NEW YORK, SPRINGER, 2021, 0175-7571

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.095

RIV identification code

RIV/00216224:14740/21:00121458

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1007/s00249-021-01497-6

UT WoS

000615765400001

Keywords in English

Protein stability; Buffer; Screening; Differential scanning fluorimetry; Dynamic light scattering; Bio-layer interferometry

Abstract

V originále

The determination of a suitable buffer environment for a protein of interest is not an easy task. 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. 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 less than half a milligram of protein in 1 day. The application of this screen helps to avoid unsuitable conditions, to explain problems observed upon protein analysis and to choose the most suitable buffers for further research. The screen can be routinely used as a primary screen for buffer optimization in labs and facilities.

Links

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LM2018131, research and development project

Name: Česká národní infrastruktura pro biologická data (Acronym: ELIXIR-CZ)

Investor: Ministry of Education, Youth and Sports of the CR, Czech National Infrastructure for Biological Data

Citovat

HOUSER, Josef, Jana KOSOUROVÁ, Monika KUBÍČKOVÁ and Michaela WIMMEROVÁ. Development of 48-condition buffer screen for protein stability assessment. European Biophysics Journal With Biophysics Letters. NEW YORK: SPRINGER, 2021, vol. 50, 3-4, p. 461-471. ISSN 0175-7571. Available from: https://dx.doi.org/10.1007/s00249-021-01497-6.

@article{1762616,
   author = {Houser, Josef and Kosourová, Jana and Kubíčková, Monika and Wimmerová, Michaela},
   article_location = {NEW YORK},
   article_number = {3-4},
   doi = {http://dx.doi.org/10.1007/s00249-021-01497-6},
   keywords = {Protein stability; Buffer; Screening; Differential scanning fluorimetry; Dynamic light scattering; Bio-layer interferometry},
   language = {eng},
   issn = {0175-7571},
   journal = {European Biophysics Journal With Biophysics Letters},
   title = {Development of 48-condition buffer screen for protein stability assessment},
   url = {https://link.springer.com/article/10.1007%2Fs00249-021-01497-6},
   volume = {50},
   year = {2021}
}

TY  - JOUR
ID  - 1762616
AU  - Houser, Josef - Kosourová, Jana - Kubíčková, Monika - Wimmerová, Michaela
PY  - 2021
TI  - Development of 48-condition buffer screen for protein stability assessment
JF  - European Biophysics Journal With Biophysics Letters
VL  - 50
IS  - 3-4
SP  - 461-471
EP  - 461-471
PB  - SPRINGER
SN  - 01757571
KW  - Protein stability
KW  - Buffer
KW  - Screening
KW  - Differential scanning fluorimetry
KW  - Dynamic light scattering
KW  - Bio-layer interferometry
UR  - https://link.springer.com/article/10.1007%2Fs00249-021-01497-6
N2  - The determination of a suitable buffer environment for a protein of interest is not an easy task. 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. 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 less than half a milligram of protein in 1 day. The application of this screen helps to avoid unsuitable conditions, to explain problems observed upon protein analysis and to choose the most suitable buffers for further research. The screen can be routinely used as a primary screen for buffer optimization in labs and facilities.
ER  -

HOUSER, Josef, Jana KOSOUROVÁ, Monika KUBÍČKOVÁ and Michaela WIMMEROVÁ. Development of 48-condition buffer screen for protein stability assessment. \textit{European Biophysics Journal With Biophysics Letters}. NEW YORK: SPRINGER, 2021, vol.~50, 3-4, p.~461-471. ISSN~0175-7571. Available from: https://dx.doi.org/10.1007/s00249-021-01497-6.

Detailed Information on Publication Record