Stability of Protein Formulations at Subzero Temperatures by Isochoric Cooling

Correia, Cátia; Tavares, Evandro; Lopes, Carlos; Silva, Joana G.; Duarte, Andreia; Rodrigues, Miguel A.; Melo, Eduardo P.

doi:10.1016/j.xphs.2019.06.017

Cited by 15 publications

(12 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Due to almost negligible material cost, it could be suitable for routine deployment in a variety of related process monitoring contexts e.g. isochoric freezing of pharmaceuticals in vials; [53][54] for the immediate needs of SPRF, it should allow for implementation of postselection strategies to diminish the large statistical variations we witnessed (FIG. 2 and FIG.…”

Section: Discussionmentioning

confidence: 99%

Self-pressurized rapid freezing at arbitrary cryoprotectant concentrations

Rolle

Okotrub

Babin

et al. 2022

Preprint

View full text Add to dashboard Cite

Self-pressurized rapid freezing (SPRF) has been proposed as a simple alternative to traditional high pressure freezing (HPF) protocols for vitrification of biological samples in electron microscopy and cryopreservation applications. Both methods exploit the circumstance that the melting point of ice reaches a minimum when subjected to pressure of around 210 [MPa], however, in SPRF its precise quantity depends on sample properties and hence, is generally unknown. In particular, cryoprotective agents (CPAs) are expected to be a factor; though eschewed by many SPRF experiments, vitrification of larger samples notably cannot be envisaged without them. Thus, in this study, we address the question of how CPA concentration affects pressure inside sealed capillaries, and how to design SPRF experiments accordingly. By embedding a fiber-optic probe in samples and performing Raman spectroscopy after freezing, we first present a direct assessment of pressure buildup during SPRF, enabled by the large pressure sensitivity of the Raman shift of hexagonal ice. Choosing dimethyl sulfoxide (DMSO) as a model CPA, this approach allows us to demonstrate that average pressure drops to zero when DMSO concentrations of 15 wt % are exceeded. Since a trade-off between pressure and DMSO concentration represents an impasse with regards to vitrification of larger samples, we introduce a sample architecture with two chambers, separated by a partition that allows for equilibration of pressure but not DMSO concentrations. We show that pressure and concentration in the fiber-facing chamber can be tuned independently, and present differential scanning calorimetry (DSC) data supporting the improved vitrification performance of two-chamber designs.

show abstract

Section: Discussionmentioning

confidence: 99%

Self-pressurized rapid freezing at arbitrary cryoprotectant concentrations

Rolle

Okotrub

Babin

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Data were placed on an absolute scale by normalizing the intensity to the incident beam flux. Analyses of the radially averaged intensities were performed in the Guinier regime ( Q R g < 1.3) by using eq , where R g is the apparent value of the radius of gyration and I (0) is the scattered intensity at zero scattering angle. …”

Section: Methodsmentioning

confidence: 99%

High-Pressure, Low-Temperature Induced Unfolding and Aggregation of Monoclonal Antibodies: Role of the Fc and Fab Fragments

et al. 2022

View full text Add to dashboard Cite

The effects of high pressure and low temperature on the stability of two different monoclonal antibodies (MAbs) were examined in this work. Fluorescence and small-angle neutron scattering were used to monitor the in situ effects of pressure to infer shifts in tertiary structure and characterize aggregation prone intermediates. Partial unfolding was observed for both MAbs, to different extents, under a range of pressure/temperature conditions. Fourier transform infrared spectroscopy was also used to monitor ex situ changes in secondary structure. Preservation of native secondary structure after incubation at elevated pressures and subzero ° C temperatures was independent of the extent of tertiary unfolding and reversibility. Several combinations of pressure and temperature were also used to discern the respective contributions of the isolated Ab fragments (Fab and Fc) to unfolding and aggregation. The fragments for each antibody showed significantly different partial unfolding profiles and reversibility. There was not a simple correlation between stability of the full MAb and either the Fc or Fab fragment stabilities across all cases, demonstrating a complex relationship to full MAb unfolding and aggregation behavior. That notwithstanding, the combined use of spectroscopic and scattering techniques provides insights into MAb conformational stability and hysteresis in high-pressure, low-temperature environments.

show abstract

“…Sucrose, glycerol, and L-arginine are three osmolytes that greatly enhanced the stability of protein disulfide isomerase A1 at −20°C, with all three tested under isoelectric focusing (Correia et al, 2020).…”

Section: Appli C Ati On Approachmentioning

confidence: 99%

“…Different solutions (sucrose, glycerol, and L‐arginine) through osmotic methods greatly enhanced the durability of binding protein isomerase A1 at −20°C, as indicated by isochoric cooling over a 700‐h period. Insulin at 20°C with isochoric cooling decreased 22% of its activity after 15 days, but 0.6 M sucrose solution limits insulin degradation (Correia et al, 2020).…”

Section: Isochoric Influence On Food Macromoleculesmentioning

confidence: 99%

Isochoric freezing of foods: A review of instrumentation, mechanism, physicochemical influence, and applications

Thakur

Jha

Bhardwaj

et al. 2022

Food Processing Preservation

View full text Add to dashboard Cite

This literature survey reveals the extended shelf life of foods when subjected to isochoric freeze processing. All other preservation aspects, thermal or non‐thermal, contribute to food preservation as per their specific functionality. From the given set of available food preservation technologies, shelf life extension by way of isochoric freezing works very well to extend the shelf life of fresh agricultural produce, without any weight loss and browning reaction to occur. Similarly, isochoric freezing of spinach tends to retain the amount of ascorbic acid and phenolic compounds in it. On the other hand, preservation by conventional freezing results in weight loss, flavor, and odor degradation, for example in fish. The isochoric method act as panacea for such types of problems faced during storage. This method works on to the key element wherein maintenance of intracellular osmolarity prevent the passing of solute concentration gradients across bind and free space inside the food. Practical Applications This emerging novel processing of foods is applicable for fruits and vegetables to preserve them with minimal fresh harvest produce loss. Isochoric processing is done at a sub‐zero temperature, which keeps the food fresh longer. This sustainability will lead to the minimization of price fluctuation and will also addresses food safety and security, minimize post‐harvest losses, and enhance market appealability. This process has also revealed its potential among poultry and other meat products with enhanced keeping qualities.

show abstract

Stability of Protein Formulations at Subzero Temperatures by Isochoric Cooling

Cited by 15 publications

References 34 publications

Self-pressurized rapid freezing at arbitrary cryoprotectant concentrations

Self-pressurized rapid freezing at arbitrary cryoprotectant concentrations

High-Pressure, Low-Temperature Induced Unfolding and Aggregation of Monoclonal Antibodies: Role of the Fc and Fab Fragments

Isochoric freezing of foods: A review of instrumentation, mechanism, physicochemical influence, and applications

Contact Info

Product

Resources

About