4.5 Article

Effect of Aluminum Adjuvant and Preservatives on Structural Integrity and Physicochemical Stability Profiles of Three Recombinant Subunit Rotavirus Vaccine Antigens

Journal

JOURNAL OF PHARMACEUTICAL SCIENCES
Volume 109, Issue 1, Pages 476-487

Publisher

ELSEVIER SCIENCE INC
DOI: 10.1016/j.xphs.2019.10.004

Keywords

rotavirus; vaccine; adjuvant; formulation; stability; thimerosal; preservative

Funding

  1. Bill and Melinda Gates Foundation

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A nonreplicating rotavirus vaccine (NRRV) containing 3 recombinant fusion proteins adsorbed to aluminum adjuvant (Alhydrogel [AH]) is currently in clinical trials. The compatibility and stability of monovalent NRRV antigen with key components of a multidose vaccine formulation were examined using physicochemical and immunochemical methods. The extent and strength of antigen-adjuvant binding were diminished by increasing phosphate concentration, and acceptable levels were identified along with alternate buffering agents. Addition of the preservative thimerosal destabilized AH-adsorbed P2-VP8-P[8] as measured by differential scanning calorimetry. Over 3 months at 4 degrees C, AH-adsorbed P2-VP8-P[8] was stable, whereas at 25 degrees C and 37 degrees C, instability was observed which was greatly accelerated by thimerosal addition. Loss of antibody binding (enzyme-linked immunosorbent assay) correlated with loss of structural integrity (differential scanning calorimetry, fluorescence spectroscopy) with concomitant nonnative disulfide bond formation (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and Asn deamidation (liquid chromatography-mass spectrometry peptide mapping). An alternative preservative (2-phenoxyethanol) showed similar antigen destabilization. Due to limited availability, only key assays were performed with monovalent P2-VP8-P[4] and P2-VP8-P[6] AH-adsorbed antigens, and varying levels of preservative incompatibility were observed. In summary, monovalent AH-adsorbed NRRV antigens stored at 4 degrees C showed good stability without preservatives; however, future formulation development efforts are required to prepare a stable, preservative-containing, multidose NRRV formulation. (C) 2020 The Authors. Published by Elsevier Inc. on behalf of the American Pharmacists Association (R). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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