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Evaluation of Predictors of Protein Relative Stability Obtained by Solid-State Hydrogen/Deuterium Exchange Monitored by FTIR

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A Correction to this article was published on 17 February 2021

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Abstract

Purpose

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 (Tg). The purpose of this study was to determine whether solid-state protein stability correlates with the difference between storage temperature and apparent Td 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. H/D exchange of freeze-dried formulations at 11% RH and temperatures from 30 to 80°C was monitored. Protein stability against aggregation at 40°C/11% RH for 6 months was assessed by size exclusion chromatography (SEC).

Results

Both sucrose and trehalose showed equivalent protection of protein secondary structure by FTIR. The rHSA:sucrose formulation showed superior long-term stability at 40°C by SEC over the trehalose formulation, but the apparent Td determined from H/D exchange was much higher in the trehalose formulation. Instead, the extent of H/D exchange (X∞) was lower in the sucrose formulation at the temperature of the stability studies (40°C) than found for the trehalose formulation, which was consistent with better stability in the sucrose formulation.

Conclusions

While apparent Td did not correlate with protein stability for rHSA, the extent of H/D exchange, X∞, did.

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Author notes

  1. Rui Fang

    Present address: Sterile and Specialty Products, Merck & Co., Inc, NJ, Kenilworth, USA

Authors and Affiliations

  1. University of Connecticut, Storrs, Connecticut, USA

    Rui Fang, Wasfy Obeidat, Michael J. Pikal & Robin H. Bogner

  2. Department of Pharmaceutical Technology, Faculty of Pharmacy, Jordan University of Science and Technology, Irbid, Jordan

    Wasfy Obeidat

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  1. Rui Fang
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  2. Wasfy Obeidat
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  4. Robin H. Bogner
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Correspondence to Robin H. Bogner.

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The original online version of this article was revised to add the following affiliation as a second affiliation to Dr. Wasfy Obeidat. The second affiliation is Department of Pharmaceutical Technology, Faculty of Pharmacy, Jordan University of Science and Technology, Irbid, Jordan.

In Loving Memory of Professor Michael J. Pikal

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Fang, R., Obeidat, W., Pikal, M.J. et al. Evaluation of Predictors of Protein Relative Stability Obtained by Solid-State Hydrogen/Deuterium Exchange Monitored by FTIR. Pharm Res 37, 168 (2020). https://doi.org/10.1007/s11095-020-02897-7

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  • DOI: https://doi.org/10.1007/s11095-020-02897-7

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