In-line near infrared spectroscopy during freeze-drying as a tool to measure efficiency of hydrogen bond formation between protein and sugar, predictive of protein storage stability

Mensink, Maarten A; Bockstal, Pieter-Jan Van; Pieters, Sigrid; Meyer, Laurens De; Frijlink, Henderik W.; Maarschalk, Kees van der Voort; Hinrichs, Wouter L. J.; Beer, Thomas De

doi:10.1016/j.ijpharm.2015.11.030

Cited by 35 publications

(29 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previously, we reported that smaller and molecularly more flexible sugars were better capable of stabilizing proteins during lyophilization (11), as they were less inhibited by steric hindrance and configurational inflexibility and therewith better able to form hydrogen bonds with the protein during lyophilization (12). Here, it was found that miscibility and absolute 1 H T 1 relaxation times decreased with increasing molecular weight of the sugar used.…”

Section: Discussionsupporting

confidence: 49%

“…Recently, it was shown that larger and molecularly more rigid sugars (e.g., oligo-and polysaccharides with a backbone through the sugar ring) are less efficient stabilizers of proteins than smaller sugars (e.g., disaccharides) (11). It was found that these larger sugars form fewer hydrogen bonds with the protein during the last part of lyophilization (12). This lack of interactions could be responsible for phase separation in these protein-sugar mixtures.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Influence of Miscibility of Protein-Sugar Lyophilizates on Their Storage Stability

Mensink

Nethercott

Hinrichs

et al. 2016

AAPS J

Self Cite

View full text Add to dashboard Cite

For sugars to act as successful stabilizers of proteins during lyophilization and subsequent storage, they need to have several characteristics. One of them is that they need to be able to form interactions with the protein and for that miscibility is essential. To evaluate the influence of protein-sugar miscibility on protein storage stability, model protein IgG was lyophilized in the presence of various sugars of different molecular weight. By comparing solid-state nuclear magnetic resonance spectroscopy relaxation times of both protein and sugar on two different timescales, i.e., (1)H T1 and (1)H T1ρ, miscibility of the two components was established on a 2-5- and a 20-50-nm length scale, respectively, and related to protein storage stability. Smaller sugars showed better miscibility with IgG, and the tendency of IgG to aggregate during storage was lower for smaller sugars. The largest sugar performed worst and was phase separated on both length scales. Additionally, shorter protein (1)H T1 relaxation times correlated with higher aggregation rates during storage. The enzyme-linked immunosorbent assay (ELISA) assay showed overlapping effects of aggregation and chemical degradation and did not correspond as well with the miscibility. Because of the small scale at which miscibility was determined (2-5 nm) and the size of the protein domains (∼2.5 × 2.5 × 5 nm), the miscibility data give an indirect measure of interaction between protein and sugar. This reduced interaction could be the result of steric hindrance, providing a possible explanation as to why smaller sugars show better miscibility and storage stability with the protein.

show abstract

Section: Discussionsupporting

confidence: 49%

Section: Introductionmentioning

confidence: 99%

Influence of Miscibility of Protein-Sugar Lyophilizates on Their Storage Stability

Mensink

Nethercott

Hinrichs

et al. 2016

AAPS J

Self Cite

View full text Add to dashboard Cite

show abstract

“… 23 This was explained in terms of the fact that the smaller and more flexible carbohydrates are less affected by steric hindrance and therefore better suited to form hydrogen bonds with the protein. 6 In this work we examine freeze-dried protein–carbohydrate samples using carbohydrates of different sizes and molecular flexibility and evaluate the ability of FTIR and THz-TDS to measure protein–carbohydrate hydrogen bonding and thermally induced changes in mobility in the amorphous state. In particular, a small disaccharide (trehalose), a molecularly flexible oligosaccharide (inulin), and a molecularly rigid polysaccharide (dextran) are used in combination with an elaborately studied protein (BSA).…”

Section: Introductionmentioning

confidence: 99%

Thermal Gradient Mid- and Far-Infrared Spectroscopy as Tools for Characterization of Protein Carbohydrate Lyophilizates

et al. 2017

Self Cite

View full text Add to dashboard Cite

Protein drugs play an important role in modern day medicine. Typically, these proteins are formulated as liquids requiring cold chain processing. To circumvent the cold chain and achieve better storage stability, these proteins can be dried in the presence of carbohydrates. We demonstrate that thermal gradient mid- and far-infrared spectroscopy (FTIR and THz-TDS, respectively) can provide useful information about solid-state protein carbohydrate formulations regarding mobility and intermolecular interactions. A model protein (BSA) was lyophilized in the presence of three carbohydrates with different size and protein stabilizing capacity. A gradual increase in mobility was observed with increasing temperature in formulations containing protein and/or larger carbohydrates (oligo- or polysaccharides), lacking a clear onset of fast mobility as was observed for smaller molecules. Furthermore, both techniques are able to identify the glass transition temperatures (Tg) of the samples. FTIR provides additional information as it can independently monitor changes in protein and carbohydrate bands at the Tg. Lastly, THz-TDS confirms previous findings that protein–carbohydrate interactions decrease with increasing molecular weight of the carbohydrate, which results in decreased protein stabilization.

show abstract

“…Moreover, many long chain carbohydrates have biopreserving properties, mostly due to their high glass transition temperature (T g ) rather than to their direct interaction with an embedded biostructure, less effective than in disaccharides due to their steric hindrance and large free volume [100][101][102]. However, notwithstanding their high T g , the polysaccharides efficiency works only up to the complete vitrification of the system: a T g much higher than the useful interval of temperatures does not improve the system protective ability [103].…”

Section: Saccharide Mixed With Other Bio-compatible Moleculesmentioning

confidence: 99%

“…The presence of small components within the solvent part of the matrix enhances the ability of the whole matrix to interact with the embedded protein, resulting in a synergistic protection [101,105], in particular if the small component is able to bind directly to a significant extent [98,106]. Indeed, adding a small interacting component helps to reduce the residual mobility of the protein even in the solid state, improving the global system efficiency [103]. The presence of small saccharides can also have a strengthening effect on the polysaccharide fraction, producing more stable networks both acting by cross-linking the polymer chains and by reducing the amount of free water around the polysaccharide [107].…”

Section: Saccharide Mixed With Other Bio-compatible Moleculesmentioning

confidence: 99%

More than a Confinement: “Soft” and “Hard” Enzyme Entrapment Modulates Biological Catalyst Function

et al. 2019

View full text Add to dashboard Cite

Catalysis makes chemical and biochemical reactions kinetically accessible. From a technological point of view, organic, inorganic, and biochemical catalysis is relevant for several applications, from industrial synthesis to biomedical, material, and food sciences. A heterogeneous catalyst, i.e., a catalyst confined in a different phase with respect to the reagents’ phase, requires either its physical confinement in an immobilization matrix or its physical adsorption on a surface. In this review, we will focus on the immobilization of biological catalysts, i.e., enzymes, by comparing hard and soft immobilization matrices and their effect on the modulation of the catalysts’ function. Indeed, unlike smaller molecules, the catalytic activity of protein catalysts depends on their structure, conformation, local environment, and dynamics, properties that can be strongly affected by the immobilization matrices, which, therefore, not only provide physical confinement, but also modulate catalysis.

show abstract

In-line near infrared spectroscopy during freeze-drying as a tool to measure efficiency of hydrogen bond formation between protein and sugar, predictive of protein storage stability

Cited by 35 publications

References 29 publications

Influence of Miscibility of Protein-Sugar Lyophilizates on Their Storage Stability

Influence of Miscibility of Protein-Sugar Lyophilizates on Their Storage Stability

Thermal Gradient Mid- and Far-Infrared Spectroscopy as Tools for Characterization of Protein Carbohydrate Lyophilizates

More than a Confinement: “Soft” and “Hard” Enzyme Entrapment Modulates Biological Catalyst Function

Contact Info

Product

Resources

About