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Rapid Identification of Pathogens01:25

Rapid Identification of Pathogens

MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...

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Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses
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An HPLC-SEC-based rapid quantification method for vesicular stomatitis virus particles to facilitate process

Adrian Schimek1, Judy K M Ng1, Ioannes Basbas1

  • 1ViraTherapeutics GmbH, Bundesstraße 27, 6063 Rum, Austria.

Molecular Therapy. Methods & Clinical Development
|May 22, 2024
PubMed
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A new label-free method accurately quantifies enveloped virus particles (VPs) using size-exclusion chromatography (SEC) and HPLC. This efficient technique is crucial for developing virus-based therapies and ensuring process consistency.

Keywords:
HPLCVSV-GPanalytical SECvesicular stomatitis virusvirus particle quantification

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Process Development

Background:

  • Accurate virus particle (VP) quantification is essential for producing VPs for therapeutic applications.
  • Current methods may lack efficiency or require labeling, hindering process development.

Purpose of the Study:

  • To develop and characterize a label-free method for quantifying enveloped VPs.
  • To establish a reliable process performance indicator for VP production.

Main Methods:

  • Utilized size-exclusion chromatography (SEC) coupled with high-performance liquid chromatography (HPLC).
  • Employed ultraviolet (UV) detection for quantification and multi-angle light scattering (MALS) for characterization.
  • Optimized mobile phase and used bovine serum albumin (BSA) for sample stabilization, achieving >97% recovery.

Main Results:

  • Demonstrated high precision with <1% repeatability and <3% intermediate precision variation.
  • Established a wide linear range (7.08 × 10^8 to 1.72 × 10^11 VP/mL), with LOD of 7.72 × 10^7 VP/mL and LLOQ of 4.20 × 10^8 VP/mL.
  • Validated applicability to in-process samples.

Conclusions:

  • The developed SEC-HPLC method offers a precise, efficient, and label-free approach for VP quantification.
  • This method supports process development for virus-based therapies, including oncolytic virotherapy, vaccine development, and gene therapy.
  • The assay's speed and minimal hands-on time facilitate same-day results for process monitoring.