Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

High Resolution Removal of Virus from Protein Solutions Using a Membrane of Unique Structure

Bio/Technology volume 10, pages 182–188 (1992)Cite this article

Abstract

We describe a new class of membrane that has the capability of removing particles such as viruses from solution with resolution and reproducibility superior to that of conventional membranes. This composite membrane is composed of a pre-formed microporous membrane plus a thin asymmetric, finely porous retentive layer that is quite different from conventional ultrafilters. The protein sieving characteristics of this membrane are nearly equivalent to, but slightly less than, that of conventional 100,000 Dalton cut-off ultrafiltration membranes. This membrane uniquely shows particle retention characteristics that increase monotonically from 3 to 8 logs as a function of particle diameter in the range of 28 to 93 nm. The performance of this membrane in both a single stage and a two stage system show that 4 to 6 log overall removal of virus particles in the size range 30 to 70 nm is possible with simultaneous high recovery of product protein. Clearance factors exceeding 6 logs are possible with viruses larger than 78 nm. In addition, the performance of process systems containing this membrane is predictable in accordance with the general membrane properties and equilibrium mass balance models. This membrane system is fully validatable and can be used in conjunction with other validated operations in a downstream process to reliably achieve an overall reduction of 12 logs of known or putative virus particles.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Lubiniecki, A.S. 1988. Safety of recombinant biologics: Issues and emerging answers, p. 3: 3–12. In: Animal Cell Biotechnology, Spier, R.E. and Griffiths, J.B. (Eds.). Academic Press, London, UK.

    Google Scholar 

  2. Petricciani, J.C. 1985. Regulatory considerations for products derived from the new biotechnology. Pharmaceut. Manufac. 5: 31–34.

    Google Scholar 

  3. Builder, S., van Reis, R., Paoni, N., Ogez, J. 1989. Process development and regulatory approval of tissue-type plasminogen activator, In: Advances in Animal Cell Biology and Technology for Bioprocesses. Spier, R.E., Griffiths, J.B., Stephenne, J., Crooy, P.J. (Eds.). Butterworths, Stoneham, MA. 452–464.

    Google Scholar 

  4. Division of Blood and Blood Products, U. S. Food and Drug Administration. 1984. Points to consider in the characterization of cell lines used to produce biologicals.

  5. Office of Biologicals Research and Review, U. S. Food and Drug Administration. 1985. Points to consider in the production and testing of new drugs and biologicals by recombinant DNA technology.

  6. Eshkol, A., Maillard, F. and Stiles, G. 1988. Purification of biological fluid by filtration through ultrafiltration membrane. European Patent No. EP0307373A2.

  7. Wiebe, M., Becker, F., Lazar, L., May, L., Casto, B., Semense, M., Fautz, C., Garnick, C., Miller, C., Masover, G., Bergmann, D. and Lubiniecki, A.S. 1989. A multifaceted approach to assure that recombinant tPA is free of adventitious virus, p. 68–71. In: Advances in Animal Cell Biology and Technology for Bioprocesses. Op. cit.

    Google Scholar 

  8. Lubiniecki, A.L. and May, L.H. 1984. Cell bank characterization for recombinant DNA mammalian cell lines. Develop. Biol. Standard. 60: 141–146.

    Google Scholar 

  9. Pocchiari, M., Macchi, G., Peano, S. and Conz, A. 1988. Can potential hazard of Creutzfeld-Jacob disease infectivity be reduced in the production of human growth hormone. Arch. Virol. 98: 131–135.

    Article  CAS  Google Scholar 

  10. Bechtel, M. K., Bagdasarian, A., Olson, W.P. and Estep, T.N. 1988. Virus removal or inactivation in hemoglobin solutions by ultrafiltration or detergent/solvent treatment. Biomat., Art. Cells, Art. Org. 16: 123–128.

    Article  CAS  Google Scholar 

  11. Horowitz, B., Wiebe, M.E., Lippin, A. and Stryker, M.H. 1985 Inactivation of viruses in labile blood derivatives. Transfusion 25: 516–522.

    Article  CAS  Google Scholar 

  12. Microbiological Associates, Inc. 1988. Evaluation of removal of simian virus 40 from human growth hormone by PTHK membrane purification process, In: Validation of Ares Serono Pharmaceutical Production, Ares-Serono S.A. Switzerland. 1: 41.

  13. Prince, A.M., Stephen, W. and Brotman, B. 1984. Beta-propiolactone/UV irradiation: A review of its effectiveness Rev. of Inf. Diseases. 5: 92–107.

  14. HyClone Laboratories, Inc. 1989. Greater retention of contaminants in fetal bovine serum: 40 nm filtration. Art to Science 2: 1–4.

  15. Zeman, L. and Wales, M. 1981. Polymer solute rejection by ultrafiltration membranes, In: Synthetic Membranes, Vol II. A. F. Turbak (Ed.) Am. Chem. Soc., Washington, DC. 2: 411.

    Chapter  Google Scholar 

  16. Kassotis, J.S., Shmidt, J., Hodgins, L.T. and Gregor, H.P. 1985. Modelling of the pore size distribution of ultrafiltration membranes. J. Memb. Sci. 22: 61–76.

    Article  CAS  Google Scholar 

  17. Steuck, M.J. Porous membrane having hydrophilic surface and process. US Patent 4,618,533.

  18. DiLeo, A.J., Allegrezza, A.E. Jr. and Burke, E.T. 1991. Membrane, process and system for isolating virus from solution. US Patent 5017292.

  19. Pitt, A. 1987. The non-specific binding of polymeric microporous membranes. J. Parenteral Sci. and Techn. 41: 110–113.

    CAS  Google Scholar 

  20. Fasman, G. (Ed.). 1976. Handbook of Biochemistry and Molecular Biology, 3rd Ed. CRC Press, Boca Raton, FL. 242

    Google Scholar 

  21. Bird, R.B., Stewart, W.E. and Lightfoot, E.N., 1960. Transport Phenomena, John Wiley and Sons, Inc., NY. 514.

  22. Millipore Corporation. 1988. An introduction to protein processing by tangential flow filtration. Process Systems Technical Brief, TB032.

  23. McCabe, W.L. and Smith, J.C. 1967. Equilibrium staged operations, In: Unit Operations of Chemical Engineering, Second Ed. McGraw-Hill Book Co., NY, NY. 528–530.

    Google Scholar 

  24. Merck Index, Tenth Ed., Martha Windholz Ed. Merck & Co., Inc., Rahway, NJ.1239.

  25. Willingmann, P., Krishnaswamy, S., McKenna, R., Smith, T.J., Olson, N.H., Rossmann, M.G., Stow, P.L. and Incardona, N.L. 1983. Preliminary investigation of the phage φX174 crystal structure. J. Mol. Biol. 212: 345–350. 1990.

    Article  Google Scholar 

  26. Vidaver, A.K., Koski, R.K. and van Etten, J.L. 1973. Bacteriophage φ6: A lipid-containing virus of Pseudomonas phaseolicola J. Virology 11: 799–805.

    CAS  PubMed  Google Scholar 

  27. Ellis, L.F. and Schlegel, R.A. 1974. Electron microscopy of Pseudomonas φ6 bacteriophage. J. Virology 14: 1547–1551.

    CAS  PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Anthony J. DiLeo: Corresponding author.

Authors and Affiliations

  1. Millipore Corporation, 80 Ashby Rd., Bedford, MA, 01730

    Anthony J. DiLeo & Anthony E. Allegrezza Jr.

  2. Genentech, Inc., 460 Point San Bruno Blvd., South San Francisco, Ca, 94080

    Stuart E. Builder

Authors
  1. Anthony J. DiLeo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anthony E. Allegrezza Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stuart E. Builder
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

DiLeo, A., Allegrezza, A. & Builder, S. High Resolution Removal of Virus from Protein Solutions Using a Membrane of Unique Structure. Nat Biotechnol 10, 182–188 (1992). https://doi.org/10.1038/nbt0292-182

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0292-182

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing