Lysozyme-Sucrose Interactions in the Solid State: Glass Transition, Denaturation, and the Effect of Residual Water

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Lysozyme-Sucrose Interactions in the Solid State : Glass Transition, Denaturation, and the Effect of Residual Water

Bogdanova, Ekaterina (author): Malmö University,Malmö universitet,Institutionen för biomedicinsk vetenskap (BMV),Biofilms Research Center for Biointerfaces

Lages, Sebastian (author): Malmö University,Lund University,Lunds universitet,Malmö universitet,Institutionen för biomedicinsk vetenskap (BMV),Biofilms Research Center for Biointerfaces,MAX IV Laboratory, Lund University, Lund SE-22484, Sweden,MAX IV-laboratoriet

Phan-Xuan, Tuan (author): Malmö University,Lund University,Lunds universitet,Malmö universitet,Institutionen för biomedicinsk vetenskap (BMV),Biofilms Research Center for Biointerfaces,MAX IV Laboratory, Lund University, Lund SE-22484, Sweden,MAX IV-laboratoriet

Kamal, Md Arif (author): Malmö University,Malmö universitet,Institutionen för biomedicinsk vetenskap (BMV),Biofilms Research Center for Biointerfaces,Division of Physical Chemistry, Lund University, Box 124, Lund SE-221 00, Sweden

Terry, Ann (author): Lund University,Lunds universitet,MAX IV-laboratoriet,MAX IV Laboratory

Millqvist-Fureby, Anna (author): RISE,Kemiska processer och läkemedel,RISE Research Institutes of Sweden, Stockholm SE-114 86, Sweden

Kocherbitov, Vitaly (author): Malmö University,Malmö universitet,Institutionen för biomedicinsk vetenskap (BMV),Biofilms Research Center for Biointerfaces

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American Chemical Society, 2023
2023
English.
In: Molecular Pharmaceutics. - : American Chemical Society. - 1543-8384 .- 1543-8392. ; 20:9, s. 4664-

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Journal article (peer-reviewed)

Abstract Subject headings

The freeze-drying of proteins, along with excipients, offers a solution for increasing the shelf-life of protein pharmaceuticals. Using differential scanning calorimetry, thermogravimetric analysis, sorption calorimetry, and synchrotron small-angle X-ray scattering (SAXS), we have characterized the properties at low (re)hydration levels of the protein lysozyme, which was freeze-dried together with the excipient sucrose. We observe that the residual moisture content in these samples increases with the addition of lysozyme. This results from an increase in equilibrium water content with lysozyme concentration at constant water activity. Furthermore, we also observed an increase in the glass transition temperature (Tg) of the mixtures with increasing lysozyme concentration. Analysis of the heat capacity step of the mixtures indicates that lysozyme does not participate in the glass transition of the sucrose matrix; as a result, the observed increase in the Tg of the mixtures is the consequence of the confinement of the amorphous sucrose domains in the interstitial space between the lysozyme molecules. Sorption calorimetry experiments demonstrate that the hydration behavior of this formulation is similar to that of the pure amorphous sucrose, while the presence of lysozyme only shifts the sucrose transitions. SAXS analysis of amorphous lysozyme-sucrose mixtures and unfolding of lysozyme in this environment show that prior to unfolding, the size and shape of lysozyme in a solid sucrose matrix are consistent with its native state in an aqueous solution. The results obtained from our study will provide a better understanding of the low hydration behavior of protein-excipient mixtures and support the improved formulation of biologics. © 2023 The Authors.

Subject headings

NATURVETENSKAP -- Kemi -- Fysikalisk kemi (hsv//swe)
NATURAL SCIENCES -- Chemical Sciences -- Physical Chemistry (hsv//eng)

Keyword

differential scanning calorimetry
glass transition
hydration
proteins
small-angle X-ray scattering
solid-state formulations
differential scanning calorimetry
glass transition
hydration
proteins
small-angle X-ray scattering
solid-state formulations

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By the author/editor: Bogdanova, Ekate ...; Lages, Sebastian; Phan-Xuan, Tuan; Kamal, Md Arif; Terry, Ann; Millqvist-Fureby ...; show more...; Kocherbitov, Vit ...; show less...

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NATURAL SCIENCES: NATURAL SCIENCES; and Chemical Science ...; and Physical Chemist ...

Articles in the publication: Molecular Pharma ...

By the university: RISE; Malmö University; Lund University

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