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Related Concept Videos

Western Blotting01:15

Western Blotting

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Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
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Accelerated vaccine process development by orthogonal protein characterization.

D Andrew James1, Lisa Szymkowicz1, Lois Yin1

  • 1Sanofi, Toronto, ON, Canada.

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|April 8, 2025
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Summary
This summary is machine-generated.

Developing new recombinant protein vaccines against SARS-CoV-2 variants required rapid biochemical testing. A novel suite of assays, including reversed-phase high-performance liquid-chromatography and LC/MS/MS, supported accelerated decision-making for clinical trials.

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Area of Science:

  • Biochemistry
  • Vaccinology
  • Analytical Chemistry

Background:

  • The COVID-19 pandemic necessitated accelerated vaccine development timelines.
  • Emergence of SARS-CoV-2 variants of concern (VOCs) required rapid adaptation of vaccine antigen targets.
  • Existing biochemical assays needed re-development to support new antigen variants.

Purpose of the Study:

  • To present a novel analytical method for assessing recombinant protein vaccine antigen purity and identity.
  • To demonstrate the utility of this assay suite in supporting accelerated bioprocess development and clinical trial transitions.
  • To enable rapid scientific decision-making during vaccine development under pandemic constraints.

Main Methods:

  • Development of a reversed-phase high-performance liquid-chromatography (RP-HPLC) method for antigen purity assessment.
  • Orthogonal characterization using Simple Western assays for identity verification.
  • Liquid-chromatography tandem mass spectrometry (LC/MS/MS) for comprehensive identification and structural analysis.

Main Results:

  • The RP-HPLC method provided robust assessment of antigen purity.
  • Orthogonal assays confirmed antigen identity and quantity.
  • The integrated assay suite facilitated rapid process development and characterization.
  • The assays supported a transition from Phase 2 to Phase 3 trials in under two weeks.

Conclusions:

  • A suite of orthogonal biochemical assays, including RP-HPLC, Simple Western, and LC/MS/MS, effectively supports accelerated development of recombinant protein vaccines.
  • This analytical approach enables rapid characterization and decision-making crucial for meeting urgent public health needs.
  • The presented methods are vital for adapting vaccine candidates to emerging viral variants.