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Related Concept Videos

Dialysis01:15

Dialysis

582
Dialysis is a diffusion-based purification process that separates analyte molecules from a complex matrix. This is accomplished by allowing molecules in the solution to pass through a semipermeable membrane into a liquid on the other side. The membrane is usually made of cellulose acetate or cellulose nitrate, and the second liquid must be miscible with the solution. Ions (e.g., chloride or sodium) or organic molecules (e.g., glucose) can pass through the membrane pores, which generally have...
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Updated: Jun 3, 2025

Author Spotlight: Exploring Macrophage Immunometabolism Through Lentiviral Vector-Mediated Gene Manipulation
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Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing.

Jennifer J Labisch1, Maria Evangelopoulou1,2, Tobias Schleuß3

  • 1Lab Essentials Applications Development Sartorius Göttingen Germany.

Engineering in Life Sciences
|January 13, 2025
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Summary

Optimizing lentiviral vector (LV) ultrafiltration is key for gene therapy. Reinforced PES and Hydrosart membranes in stirred cells best concentrate infectious LVs while removing DNA impurities.

Keywords:
centrifugal ultrafilterscrossflow cassettesdownstream processinglentiviral vectorsstirred cellstangential flow filtrationultrafiltration

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Area of Science:

  • Biotechnology
  • Gene Therapy
  • Bioprocessing

Background:

  • Lentiviral vectors (LVs) are crucial for ex vivo gene therapies.
  • LV production faces challenges due to vector instability and fragility.
  • Downstream processing optimization is needed to improve LV yield and quality.

Purpose of the Study:

  • To investigate the impact of membrane type and filtration device on lentiviral vector ultrafiltration.
  • To identify optimal conditions for concentrating infectious LVs and removing impurities.
  • To enhance the efficiency and product quality of LV downstream processing.

Main Methods:

  • Evaluated nine membrane materials (PES, regenerated cellulose, Hydrosart) with varying molecular weight cutoffs.
  • Tested membranes using stirred cells, centrifugal ultrafilters, and crossflow cassettes.
  • Assessed performance based on infectious LV particle retention and impurity removal (DNA).

Main Results:

  • Reinforced 100 kDa PES and 300 kDa Hydrosart membranes showed superior performance in concentrating infectious LVs and removing DNA.
  • Stirred cell operation yielded better results compared to other filtration devices.
  • Non-optimized crossflow cassette processes resulted in lower infectious LV recovery.

Conclusions:

  • Membrane material and filtration device significantly influence the efficiency of lentiviral vector ultrafiltration.
  • Specific membrane types and stirred cell configurations are recommended for improved LV concentration and purity.
  • Further optimization of crossflow systems may be necessary to enhance LV recovery.