Suppression of protein inactivation during freezing by
minimizing pH changes using ionic cryoprotectants

J 2016

Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants

KRAUSKOVÁ, Ľubica, Jitka PROCHÁZKOVÁ, Martina KLAŠKOVÁ, Lenka FILIPOVÁ, Radka CHALOUPKOVÁ et. al.

Basic information

Original name

Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants

Authors

KRAUSKOVÁ, Ľubica (703 Slovakia, belonging to the institution), Jitka PROCHÁZKOVÁ (203 Czech Republic, belonging to the institution), Martina KLAŠKOVÁ (203 Czech Republic, belonging to the institution), Lenka FILIPOVÁ (203 Czech Republic, belonging to the institution), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), Stanislav MALÝ (203 Czech Republic), Jiří DAMBORSKÝ (203 Czech Republic, belonging to the institution) and Dominik HEGER (203 Czech Republic, guarantor, belonging to the institution)

Edition

International Journal of Pharmaceutics, 2016, 0378-5173

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30104 Pharmacology and pharmacy

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL URL

Impact factor

Impact factor: 3.649

RIV identification code

RIV/00216224:14310/16:00088279

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.ijpharm.2016.05.031

UT WoS

000378949800006

Keywords in English

Protein cryopreservation; Freezing/thawing; Protein aggregation; Protein stability; Quality by design; Freezing stress; Freezing-induced pH shift

Abstract

V originále

Freezing and lyophilization are often used for stabilization of biomolecules; however, this sometimes results in partial degradation and loss of biological function in these molecules. In this study we examined the effect of freezing-induced acidity changes on denaturation of the model enzyme haloalkane dehalogenase under various experimental conditions. The effective local pH of frozen solutions is shown to be the key causal factor in protein stability. To preserve the activity of frozenthawed enzymes, acidity changes were prevented by the addition of an ionic cryoprotectant, a compound which counteracts pH changes during freezing due to selective incorporation of its ions into the ice. This approach resulted in complete recovery of enzyme activity after multiple freeze-thaw cycles. We propose the utilization of ionic cryoprotectants as a new and effective cryopreservation method in research laboratories as well as in industrial processes.

Links

GA15-12386S, research and development project

Name: Dynamika a (foto)chemie polutantů na rozhraních led/vzduch a voda/vzduch z experimentu i teorie

Investor: Czech Science Foundation

LM2011028, research and development project

Name: RECETOX ? Národní infrastruktura pro výzkum toxických látek v prostředí

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

LO1214, research and development project

Name: Centrum pro výzkum toxických látek v prostředí (Acronym: RECETOX)

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

Citovat

KRAUSKOVÁ, Ľubica, Jitka PROCHÁZKOVÁ, Martina KLAŠKOVÁ, Lenka FILIPOVÁ, Radka CHALOUPKOVÁ, Stanislav MALÝ, Jiří DAMBORSKÝ and Dominik HEGER. Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants. International Journal of Pharmaceutics. 2016, vol. 509, 1-2, p. 41-49. ISSN 0378-5173. Available from: https://dx.doi.org/10.1016/j.ijpharm.2016.05.031.

@article{1358384,
   author = {Krausková, Ľubica and Procházková, Jitka and Klašková, Martina and Filipová, Lenka and Chaloupková, Radka and Malý, Stanislav and Damborský, Jiří and Heger, Dominik},
   article_number = {1-2},
   doi = {http://dx.doi.org/10.1016/j.ijpharm.2016.05.031},
   keywords = {Protein cryopreservation; Freezing/thawing; Protein aggregation; Protein stability; Quality by design; Freezing stress; Freezing-induced pH shift},
   language = {eng},
   issn = {0378-5173},
   journal = {International Journal of Pharmaceutics},
   title = {Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants},
   url = {http://dx.doi.org/10.1016/j.ijpharm.2016.05.031},
   volume = {509},
   year = {2016}
}

TY  - JOUR
ID  - 1358384
AU  - Krausková, Ľubica - Procházková, Jitka - Klašková, Martina - Filipová, Lenka - Chaloupková, Radka - Malý, Stanislav - Damborský, Jiří - Heger, Dominik
PY  - 2016
TI  - Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants
JF  - International Journal of Pharmaceutics
VL  - 509
IS  - 1-2
SP  - 41-49
EP  - 41-49
SN  - 03785173
KW  - Protein cryopreservation
KW  - Freezing/thawing
KW  - Protein aggregation
KW  - Protein stability
KW  - Quality by design
KW  - Freezing stress
KW  - Freezing-induced pH shift
UR  - http://dx.doi.org/10.1016/j.ijpharm.2016.05.031
N2  - Freezing and lyophilization are often used for stabilization of biomolecules; however, this sometimes results in partial degradation and loss of biological function in these molecules. In this study we examined the effect of freezing-induced acidity changes on denaturation of the model enzyme haloalkane dehalogenase under various experimental conditions. The effective local pH of frozen solutions is shown to be the key causal factor in protein stability. To preserve the activity of frozenthawed enzymes, acidity changes were prevented by the addition of an ionic cryoprotectant, a compound which counteracts pH changes during freezing due to selective incorporation of its ions into the ice. This approach resulted in complete recovery of enzyme activity after multiple freeze-thaw cycles. We propose the utilization of ionic cryoprotectants as a new and effective cryopreservation method in research laboratories as well as in industrial processes.
ER  -

KRAUSKOVÁ, Ľubica, Jitka PROCHÁZKOVÁ, Martina KLAŠKOVÁ, Lenka FILIPOVÁ, Radka CHALOUPKOVÁ, Stanislav MALÝ, Jiří DAMBORSKÝ and Dominik HEGER. Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants. \textit{International Journal of Pharmaceutics}. 2016, vol.~509, 1-2, p.~41-49. ISSN~0378-5173. Available from: https://dx.doi.org/10.1016/j.ijpharm.2016.05.031.

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