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Related Concept Videos

Conjugated Proteins02:50

Conjugated Proteins

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Simple proteins and protein complexes contain only amino acids. In contrast, many other proteins, called conjugated proteins, covalently bond with non-protein moieties.
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
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Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection
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Formulation Development of a COVID-19 Recombinant Spike Protein-Based Vaccine.

Emily Xiao1, Clémentine Mirabel1, Didier Clénet2

  • 1Global Vaccine Drug Product Development, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada.

Vaccines
|August 29, 2024
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Summary

This study developed a stable SARS-CoV-2 spike protein vaccine formulation. Tween 20 prevented aggregation and conformational changes, with sucrose further enhancing thermostability for a reliable COVID-19 vaccine.

Keywords:
COVID-19HDX-MSSARS-CoV-2agitation stressspike proteinstabilitythermal stressvaccine

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Area of Science:

  • Biotechnology and Pharmaceutical Sciences
  • Vaccine Development
  • Protein Biochemistry

Background:

  • Recombinant prefusion spike proteins are key vaccine candidates for SARS-CoV-2.
  • Protein aggregation and conformational changes due to mechanical stress pose significant formulation challenges.
  • Maintaining structural integrity is crucial for vaccine efficacy and stability.

Purpose of the Study:

  • To develop a stable formulation for a recombinant prefusion spike protein vaccine against SARS-CoV-2.
  • To identify pharmaceutical excipients that prevent mechanical stress-induced aggregation and conformational changes.
  • To assess the long-term stability and thermostability of the optimized formulation.

Main Methods:

  • Screening of pharmaceutical excipients, including Tween 20, for stabilizing properties.
  • Concentration-dependent studies of Tween 20 to determine optimal levels.
  • Characterization of protein conformation using size exclusion chromatography (SEC) and hydrogen-deuterium exchange mass spectrometry (HDX-MS).
  • Long-term stability modeling and thermal shift assays (TSA) with stabilizers like sucrose and glycerol.

Main Results:

  • Tween 20 effectively inhibited mechanically induced aggregation and prevented conformational changes at 0.2% (v/v).
  • SEC and HDX-MS confirmed that Tween 20 maintained the trimeric conformation, preserving S2 domain binding.
  • Stability modeling predicted a two-year shelf life at 2-8 °C for the Tween 20 formulation.
  • Sucrose and glycerol significantly increased the spike protein's melting temperature (Tm), enhancing thermostability.

Conclusions:

  • A formulation containing 0.2% (v/v) Tween 20 at neutral pH provides adequate stability for a SARS-CoV-2 spike protein vaccine under refrigeration.
  • The addition of sucrose further improves vaccine thermostability, offering broader storage and handling options.
  • This systematic formulation approach addresses mechanical stress impacts and highlights the importance of excipients for protein vaccine integrity.