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Murine leukemia virus clearance by flocculation and microfiltration.

Saengchai Akeprathumchai1, Binbing Han, S Ranil Wickramasinghe

  • 1Department of Chemical Engineering, Colorado State University, Fort Collins, Colorado 80523-1370, USA.

Biotechnology and Bioengineering
|October 30, 2004
PubMed
Summary

Flocculation and microfiltration effectively removed amphotropic murine leukemia virus (A-MLV) from cell suspensions. Adding a flocculant was crucial for efficient virus clearance, especially with larger pore size membranes, and improved permeate flux.

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Area of Science:

  • Biotechnology
  • Virology
  • Filtration Technology

Background:

  • Murine leukemia virus (MLV) is a retrovirus used for validating retroviral particle clearance.
  • Amphotropic MLV (A-MLV) vectors, incapable of replication, were employed for biosafety.
  • A-MLV does not infect CHO cells, preventing artificial reduction in virus titer.

Purpose of the Study:

  • Investigate virus clearance from cell suspensions using flocculation and microfiltration.
  • Evaluate the impact of flocculant and membrane pore size on virus removal efficiency.
  • Assess the effect of flocculation on permeate flux.

Main Methods:

  • Used amphotropic murine leukemia virus (A-MLV) vector with enhanced green fluorescent protein (EGFP).
  • Conducted flocculation of feed streams with a cationic polyelectrolyte.

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  • Performed microfiltration using 0.1 and 0.65 micrometer pore size hollow fiber membranes.

    • Main Results:

    • 0.1 micrometer membranes achieved a 1,000-fold reduction in A-MLV titer, with or without flocculant.
    • Flocculant improved permeate flux for CHO cell feed streams with 0.1 micrometer membranes.
    • 0.65 micrometer membranes showed poor A-MLV clearance without flocculant, but similar clearance to 0.1 micrometer membranes with flocculant.

    Conclusions:

    • Tangential flow microfiltration can significantly clear A-MLV.
    • Flocculation is essential for effective virus clearance when membrane pore size exceeds virus diameter.
    • Flocculation enhances permeate flux during microfiltration.