Best practices for performing freeze dryer equipment qualification are recommended, focusing on identifying methods to quantify shelf thermal uniformity (also known as "shelf surface uniformity"), equipment capability, and performance metrics of the freeze dryer essential to the pharmaceutical Quality by Design paradigm. Specific guidelines for performing shelf temperature mapping, freeze dryer equipment limit testing (the capability curve), and condenser performance metrics have been provided. Concerning shelf temperature mapping and equipment capability measurements, the importance of paying attention to the test setup and the use of appropriate testing tools are stressed.
View Article and Find Full Text PDFPurpose: Hydrogen/deuterium (H/D) exchange over a range of temperatures suggests a protein structural/mobility transition in the solid state below the system glass transition temperature (T). The purpose of this study was to determine whether solid-state protein stability correlates with the difference between storage temperature and apparent T where an abrupt change in mobility occurs, or alternatively, the extent of H/D exchange at a single temperature correlates directly to protein stability in lyophilized solids.
Methods: Solid-state H/D exchange was monitored by FTIR spectroscopy to study the extent of exchange and the apparent transition temperature in both pure recombinant human serum albumin (rHSA) and rHSA formulated with sucrose or trehalose.
Controlling ice nucleation, at a fixed higher temperature, results in larger ice crystals, which can reduce the ice/freeze-concentrate interface area where proteins can adsorb and partially unfold. Moreover, limited work has been done to address any effects on short-term stability due to a slow ramp or long isothermal hold after the ice nucleation step. The objective was to evaluate the effect of the ice nucleation temperature and residence time in the freeze-concentrate on in-process or storage stability of representative proteins, human IgG, and recombinant human serum albumin.
View Article and Find Full Text PDFInter- and intra-batch variability in heat and mass transfer during the drying phase of lyophilization is well recognized. Heat transfer variability between individual vials in the same batch arise from both different positions in the vial array and from variations in the bottom contour of the vials, both effects contributing roughly equally to variations in the effective heat transfer coefficient of the vials, K. Both effects can be measured in the laboratory, and variations in average K values as a function of vial position in the array for lab and production can be calculated by use of the simple steady-state heat and mass transfer theory.
View Article and Find Full Text PDFThe aim of this research was to evaluate the impact of variability in ice sublimation rate (dm/dt) measurement and vial heat transfer coefficient (K) on product temperature prediction during the primary drying phase of lyophilization. The mathematical model used for primary drying uses dm/dt and K as inputs to predict product temperature. A second-generation tunable diode laser absorption spectroscopy (TDLAS)-based sensor was used to measure dm/dt.
View Article and Find Full Text PDFMolecular mobility has been traditionally invoked to explain physical and chemical stability of diverse pharmaceutical systems. Although the molecular mobility concept has been credited with creating a scientific basis for stabilization of amorphous pharmaceuticals and biopharmaceuticals, it has become increasingly clear that this approach represents only a partial description of the underlying fundamental principles. An additional mechanism is proposed herein to address 2 key questions: (1) the existence of unfrozen water (i.
View Article and Find Full Text PDFSeveral controlled ice nucleation techniques have been developed to increase the efficiency of the freeze-drying process as well as to improve the quality of pharmaceutical products. Owing to the reduction in ice surface area, these techniques have the potential to reduce the degradation of proteins labile during freezing. The objective of this study was to evaluate the effect of ice nucleation temperature on the in-process stability of lactate dehydrogenase (LDH).
View Article and Find Full Text PDFCake appearance is an important attribute of freeze-dried products, which may or may not be critical with respect to product quality (i.e., safety and efficacy).
View Article and Find Full Text PDFAlthough several mathematical models of primary drying have been developed over the years, with significant impact on the efficiency of process design, models of secondary drying have been confined to highly complex models. The simple-to-use Excel-based model developed here is, in essence, a series of steady state calculations of heat and mass transfer in the 2 halves of the dry layer where drying time is divided into a large number of time steps, where in each time step steady state conditions prevail. Water desorption isotherm and mass transfer coefficient data are required.
View Article and Find Full Text PDFThis report presents calculations of the difference between the vial heat transfer coefficient of the "edge vial" and the "center vial" at all scales. The only scale-up adjustment for center vials is for the contribution of radiation from the shelf upon which the vial sits by replacing the emissivity of the laboratory dryer shelf with the emissivity of the production dryer shelf. With edge vials, scales-up adjustments are more complex.
View Article and Find Full Text PDFLyophilized proteins are generally stored below their glass transition temperature (T) to maintain long-term stability. Some proteins in the (pure) solid state showed a distinct endotherm at a temperature well below the glass transition, designated as a pre-T endotherm. The pre-T endothermic event has been linked with a transition in protein internal mobility.
View Article and Find Full Text PDFFor 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.
View Article and Find Full Text PDFThis study investigates the effect of low levels of electrolytes on storage stability in freeze-dried sucrose-based protein formulations. Both bovine serum albumin and recombinant human serum albumin were freeze dried with sucrose and alkali halides (LiCl, NaCl, KCl, RbCl, and CsCl) at selected low levels. All formulations were stored at 50 °C and 65 °C up to 2 months and then assayed for protein aggregation.
View Article and Find Full Text PDFIn small amounts, the low molecular weight excipients-sorbitol and glycerol-have been shown to stabilize lyophilized sucrose-based protein formulations. The purpose of this study was to explore the use of amino acids as low molecular weight excipients to similarly enhance stability. Model proteins, recombinant human serum albumin and α-chymotrypsin, were formulated with sucrose in combination with one of 15 amino acid additives.
View Article and Find Full Text PDFImmunogenicity of aggregated or otherwise degraded protein delivered from depots or other biopharmaceutical products is an increasing concern, and the ability to deliver stable, active protein is of central importance. We review characterization approaches for solid protein dosage forms with respect to metrics that are intended to be predictive of protein stability against aggregation and other degradation processes. Each of these approaches is ultimately motivated by hypothetical connections between protein stability and the material property being measured.
View Article and Find Full Text PDFA confocal Raman microscopic technique was optimized to more efficiently detect amorphous-amorphous phase separation in freeze-dried protein formulations. A Renishaw Raman inVia confocal microscope was used to collect 100-200 μm line maps (2 μm step size) of freeze-dried protein-excipient formulations. At each point across the line map, the composition was evaluated from the intensity of the nonoverlapping peaks representative of each component.
View Article and Find Full Text PDFThe application of key elements of quality by design (QbD), such as risk assessment, process analytical technology, and design space, is discussed widely as it relates to freeze-drying process design and development. However, this commentary focuses on constructing the Design and Control Space, particularly for the primary drying step of the freeze-drying process. Also, practical applications and considerations of claiming a process Design Space under the QbD paradigm have been discussed.
View Article and Find Full Text PDFThe objective of this study was to assess the feasibility of developing and applying a laboratory tool that can provide three-dimensional product structural information during freeze-drying and which can accurately characterize the collapse temperature (Tc ) of pharmaceutical formulations designed for freeze-drying. A single-vial freeze dryer coupled with optical coherence tomography freeze-drying microscopy (OCT-FDM) was developed to investigate the structure and Tc of formulations in pharmaceutically relevant products containers (i.e.
View Article and Find Full Text PDFDSC thermograms of solid state pure proteins often show a distinct endotherm at a temperature far below the glass transition temperature of the system (Tg). We hypothesized this endotherm represents enthalpy recovery associated with an internal mobility transition of the protein molecule. Although the existence of an internal transition has been postulated, whether this endotherm is associated with such a transition has not previously been discussed.
View Article and Find Full Text PDFAmorphous HES/disaccharide (trehalose or sucrose) formulations, with and without added polyols (glycerol and sorbitol) and disaccharide formulations of human growth hormone (hGH), were prepared by freeze drying and characterized with particular interest in methodology for using high precision density measurements to evaluate free volume changes and a focus on comparisons between "free volume" changes obtained from analysis of density data, fast dynamics (local mobility), and PALS characterization of "free volume" hole size. Density measurements were performed using a helium gas pycnometer, and fast dynamics was characterized using incoherent neutron scattering spectrometer. Addition of sucrose and trehalose to hGH decreases free volume in the system with sucrose marginally more effective than trehalose, consistent with superior pharmaceutical stability of sucrose hGH formulations well below Tg relative to trehalose.
View Article and Find Full Text PDFThe aim of the study was to comprehensively investigate the influence of the freezing step during lyophilization on the stability of gene-delivery particles in order to better understand particle stabilization during freezing. Particle size of plasmid/linear polyethylenimine (LPEI) polyplexes at two DNA concentrations and at increasing sucrose-DNA ratios was investigated separately as a function of freezing procedure, ice-nucleation temperature, residence time of the particles in a partially frozen state, or incomplete freezing. Using a numerical model, the increase in sucrose concentration and system viscosity and corresponding bimolecular reaction rates were theoretically estimated.
View Article and Find Full Text PDFThe objective of the present work was to determine whether hydrolysis in a model lyophile was influenced by general media effects with water-changing properties of the medium or via a specific mechanism of water as a reactant. Four formulations of zoniporide and sucrose (1:10) were prepared with variable amounts of sorbitol [0%-25% (w/v) of total solids). These formulations were then equilibrated at 6% and 11% relative humidity using saturated salt solutions.
View Article and Find Full Text PDFA new type of freeze-drying microscope based upon time-domain optical coherence tomography is presented here (OCT-FDM). The microscope allows for real-time, in situ 3D imaging of pharmaceutical formulations in vials relevant for manufacturing processes with a lateral resolution of <7 μm and an axial resolution of <5 μm. Correlation of volumetric structural imaging with product temperature measured during the freeze-drying cycle allowed investigation of structural changes in the product and determination of the temperature at which the freeze-dried cake collapses.
View Article and Find Full Text PDFIn this study, changes in the local conformation of aspartame were observed in annealed lyophilized glasses by monitoring changes in the distance between two labeled sites using C-(2)H rotational-echo double-resonance (REDOR) nuclear magnetic resonance (NMR) spectroscopy. Confirmation that the REDOR experiments were producing accurate distance measurement was ensured by measuring the (13)C-(15)N distance in glycine. The experiment was further verified by measuring the REDOR dephasing curve on (13)C-(2)H methionine.
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