Nanobubbles in Reconstituted Lyophilized Formulations: Interaction With Proteins and Mechanism of Formation

Jared R. Snell, N. S.Krishna Kumar, Raj Suryanarayanan, Theodore W. Randolph

Research output: Contribution to journalArticle

Abstract

Reconstitution of lyophilized disaccharide formulations results in the formation of nanosized air bubbles that persist in suspension for weeks. If proteins are present, interactions with nanobubbles may cause loss of monomeric protein and formation of subvisible particles. The goals of this work are to determine the mechanism(s) by which nanobubbles form in reconstituted lyophilized formulations and to develop strategies for reducing nanobubble generation. We hypothesize that nanobubbles are created from nanosized gas pockets within lyophilized solids, which become bubbles when the surrounding matrix is dissolved away during reconstitution. Nanosized voids may originate from small ice crystals formed within the concentrated liquid during freezing that subsequently sublime during drying. Nanobubble concentrations are correlated with the extent of mannitol crystallization during freezing. Nanosized ice crystals, induced by the release of water during mannitol crystallization, were responsible for nanobubble formation. The presence of trehalose or sucrose, in formulations with low mannitol concentrations, inhibited excipient crystallization during lyophilization and reduced nanobubble levels following reconstitution. Our results show a correlation between nanobubble formation and concentrations of insoluble IL-1ra aggregates, suggesting that minimizing nanobubble generation may be an effective strategy for reducing protein aggregation following reconstitution.

Original languageEnglish (US)
Pages (from-to)284-292
Number of pages9
JournalJournal of Pharmaceutical Sciences
Volume109
Issue number1
DOIs
StatePublished - Jan 2020

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Mannitol
Crystallization
Ice
Freezing
Interleukin 1 Receptor Antagonist Protein
Trehalose
Proteins
Freeze Drying
Disaccharides
Excipients
Sucrose
Suspensions
Gases
Air
Water

Keywords

  • Lyophilization
  • crystallization
  • nanoparticle(s)
  • protein aggregation
  • trehalose

Cite this

Nanobubbles in Reconstituted Lyophilized Formulations : Interaction With Proteins and Mechanism of Formation. / Snell, Jared R.; Kumar, N. S.Krishna; Suryanarayanan, Raj; Randolph, Theodore W.

In: Journal of Pharmaceutical Sciences, Vol. 109, No. 1, 01.2020, p. 284-292.

Research output: Contribution to journalArticle

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