The influence of residual water on the solid-state properties of freeze-dried fibrinogen

Wahl, Verena; Leitgeb, Stefan; Laggner, Peter; Pichler, Harald; Liebminger, Andreas; Khinast, Johannes G.

doi:10.1016/j.ejpb.2015.01.006

Cited by 7 publications

(6 citation statements)

References 52 publications

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“…The dissolution rates were found to decrease with increasing specific surface, most notably in the amorphous form, in contrast to expectations from classical thermodynamics. Protein conformational changes and hydrophobic surface formation upon depletion of water could be possible causes [42,43].…”

Section: Discussionmentioning

confidence: 99%

Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders

et al. 2019

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Development of tools for direct thrombus imaging represents a key step for diagnosis and treatment of stroke. Nanoliposomal carriers of contrast agents and thrombolytics can be functionalized to target blood thrombi by small protein binders with selectivity for fibrin domains uniquely formed on insoluble fibrin. We employed a highly complex combinatorial library derived from scaffold of 46 amino acid albumin-binding domain (ABD) of streptococcal protein G, and ribosome display, to identify variants recognizing fibrin cloth in human thrombus. We constructed a recombinant target as a stretch of three identical fibrin fragments of 16 amino acid peptide of the Bβ chain fused to TolA protein. Ribosome display selection followed by large-scale Enzyme-Linked ImmunoSorbent Assay (ELISA) screening provided four protein variants preferentially binding to insoluble form of human fibrin. The most specific binder variant D7 was further modified by C-terminal FLAG/His-Tag or double His-tag for the attachment onto the surface of nanoliposomes via metallochelating bond. D7-His-nanoliposomes were tested using in vitro flow model of coronary artery and their binding to fibrin fibers was demonstrated by confocal and electron microscopy. Thus, we present here the concept of fibrin-targeted binders as a platform for functionalization of nanoliposomes in the development of advanced imaging tools and future theranostics.

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Section: Discussionmentioning

confidence: 99%

Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders

et al. 2019

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“…These differences could also be connected to the protein stability because of the formation of the glassy state upon strong dehydration. There are few studies on the structural transition in the solid state of protein using scattering techniques: for example, SANS has been carried out on dried formulations in the presence of excipients − and also SAXS. , However, no SAXS and wide-angle X-ray scattering (WAXS) studies discussing the change in protein morphology under dehydrated conditions or dehydration-induced intramolecular structural changes are found in the literature.…”

Section: Introductionmentioning

confidence: 99%

Hydration-Induced Structural Changes in the Solid State of Protein: A SAXS/WAXS Study on Lysozyme

et al. 2020

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The stability of biologically produced pharmaceuticals is the limiting factor to various applications, which can be improved by formulation in solid-state forms, mostly via lyophilization. Knowledge about the protein structure at the molecular level in the solid state and its transition upon rehydration is however scarce, and yet it most likely affects the physical and chemical stability of the biological drug. In this work, synchrotron small- and wide-angle X-ray scattering (SWAXS) are used to characterize the structure of a model protein, lysozyme, in the solid state and its structural transition upon rehydration to the liquid state. The results show that the protein undergoes distortion upon drying to adopt structures that can continuously fill the space to remove the protein–air interface that may be formed upon dehydration. Above a hydration threshold of 35 wt %, the native structure of the protein is recovered. The evolution of SWAXS peaks as a function of water content in a broad range of concentrations is discussed in relation to the structural changes in the protein. The findings presented here can be used for the design and optimization of solid-state formulations of proteins with improved stability.

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“…The powders were stored at room temperature at constant levels of relative 173 humidity (RH) of 12% (LiCl), 52% (Mg(NO 3 ) 2 *6H 2 O), 75% (NaCl ) and 93% 174 (KNO 3 ), (Rockland, 1960) Since no significant differences in the water content between batches were observed, the 180 water sorption behavior seemed to be similar for each lot. (Wahl et al, 2015a) 181…”

Section: Results and Discussion 170mentioning

confidence: 98%

“…WAXS crystallinity was calculated as the ratio of the maximum of the diffraction 339 peaks divided by the baseline height. Details including figures representing the WAXS 340 pattern can be found elsewhere (Wahl et al, 2015a(Wahl et al, , 2015b . 341…”

Section: ) 332mentioning

confidence: 99%