The importance of formulation in the successful lyophilization of influenza reference materials

Abstract

Lyophilized Influenza antigen reference reagents are a critical resource in the quality control of influenza vaccines. A standard formulation has been used successfully at NIBSC for many years however, following the unexpected occurrence of a collapsed appearance in a particular batch a study was carried out to establish the impact of the sugar concentration in the formulation using modulated differential scanning calorimetry (mDSC) and nuclear magnetic resonance spectroscopy (NMR). There was a correlation between the presence and size of the mDSC eutectic temperature events and the freeze dried appearance of the cakes, which became progressively worse with increasing amounts of sugar. NMR spectroscopy could be used to positively identify and quantify the sugars in the formulations. MDSC can rapidly predict if the freeze dried appearance will be acceptable so as to assure the successful lyophilization of influenza reference preparations.

Impact of Formulation Choices on the Freeze-Drying of an Interleukin-6 Reference Material

“…This result complements our earlier finding ( Duru et al, 2015 ), which showed that for freeze-dried influenza antigen reference preparations, NaCl crystallization was a critical factor—an increase in the amorphous content using raised levels of sucrose (above 1.4% w/w) resulted in a collapsed freeze-dried appearance. From the mDSC, it was concluded that raising the sucrose content (although usually a desirable stabilizer for freeze drying), resulted in inhibition of the crystallization of the NaCl co-formulant and that this crystallization was critical in producing an acceptable freeze-dried appearance.…”

Analytical Techniques for Structural Characterization of Proteins in Solid Pharmaceutical Forms: An Overview

“…Therefore, mDSC is particularly useful for detecting T g present with other overlapping thermal events (glass transition) [131]. Therefore, mDSC is particularly useful for detecting T g present with other overlapping thermal events (glass transition) [132], to correlate eutectic temperature (lowest possible melting temperature over all of the mixing ratios for the involved component species) events leading to cake collapse [133] and to study the effects of annealing on the thermal properties of frozen sucrose solutions [134][135][136]. It was also used as an additional method to ssHDX-MS for the analysis of the conformation of lyophilized IgG1 (Table 2) [137].…”

Characterizing Protein Structure, Dynamics and Conformation in Lyophilized Solids

“…‘Reversing’ and ‘non-reversing’ heat flow components do not refer to the reversible or irreversible nature of events; they refer to the heat capacity component and time-dependent kinetic component of total heat flow, respectively. MDSC has been used to determine Tg values for systems with small changes in heat capacity [48], to correlate eutectic temperature events to cake collapse [53] and to study the effects of annealing on the thermal properties of frozen sucrose solutions [54]. …”