Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine

Ozan S. Kumru, Soraia Saleh-Birdjandi, Lorena R. Antunez, Eddy Sayeed, David Robinson, Sjoerd van den Worm, Geoffrey S. Diemer, Wilma Perez, Patrizia Caposio, Klaus Früh, Sangeeta B. Joshi, David B. Volkin

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Live attenuated viral vaccine/vector candidates are inherently unstable and infectivity titer losses can readily occur without defining appropriate formulations, storage conditions and clinical handling practices. During initial process development of a candidate vaccine against HIV-1 using a recombinant Human Cytomegalovirus vector (rHCMV-1), large vector titer losses were observed after storage at 4 °C and after undergoing freeze-thaw. Thus, the goal of this work was to develop candidate frozen liquid formulations of rHCMV-1 with improved freeze-thaw and short-term liquid stability for potential use in early clinical trials. To this end, a virus stability screening protocol was developed including use of a rapid, in vitro cell-based immunofluorescence focus assay to quantitate viral titers. A library of ∼50 pharmaceutical excipients (from various known classes of additives) were evaluated for their effect on vector stability after freeze-thaw cycling or incubation at 4 °C for several days. Certain additives including sugars and polymers (e.g., trehalose, sucrose, sorbitol, hydrolyzed gelatin, dextran 40) as well as removal of NaCl (lower ionic strength) protected rHCMV-1 against freeze-thaw mediated losses in viral titers. Optimized solution conditions (e.g., solution pH, buffers and sugar type) slowed the rate of rHCMV-1 titer losses in the liquid state at 4 °C. After evaluating various excipient combinations, three new candidate formulations were designed and rHCMV-1 stability was benchmarked against both the currently-used and a previously reported formulation. The new candidate formulations were significantly more stable in terms of reducing rHCMV-1 titer losses after 5 freeze-thaw cycles or incubation at 4 °C for 30 days. This case study highlights the utility of semi-empirical design of frozen liquid formulations of a live viral vaccine candidate, where protection against infectivity titer losses due to freeze-thaw and short-term liquid storage are sufficient to enable more rapid initiation of early clinical trials.

Original languageEnglish (US)
Pages (from-to)6696-6706
Number of pages11
JournalVaccine
Volume37
Issue number44
DOIs
StatePublished - Oct 16 2019

Keywords

  • Cytomegalovirus
  • Excipient
  • Formulation
  • Freeze-thaw
  • HIV vaccine
  • Stability

ASJC Scopus subject areas

  • Molecular Medicine
  • Immunology and Microbiology(all)
  • veterinary(all)
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

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  • Cite this

    Kumru, O. S., Saleh-Birdjandi, S., Antunez, L. R., Sayeed, E., Robinson, D., van den Worm, S., Diemer, G. S., Perez, W., Caposio, P., Früh, K., Joshi, S. B., & Volkin, D. B. (2019). Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine. Vaccine, 37(44), 6696-6706. https://doi.org/10.1016/j.vaccine.2019.09.027