Aqueous Two-Phase Partition in Biochemical Recovery from Mammalian Cell Culture

  • Jon G. Huddleston
  • Andrew Lyddiatt


A study of the fractionation of monoclonal antibodies and other solutes in spent mammalian cell culture media has been undertaken with aqueous PEG-phosphate partition systems. Manipulation of pH and average molecular weight of polymers yielded systems capable of separating transferrin from other components. Introduction into dispersed phases of a mixture of anti-BSA Sepharose and human-IgG silica promoted selective recovery of BSA, anti-huIgG monoclonal antibody and transferrin in a “four phase”, batch system distinguished by the discrete separation of antibody product and deleterious proteolytic activity into separate aqueous phases.


Batch System Acidic Protease Bottom Phase Retention Coefficient Partition System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    H. Schutte, K.H. Kroner, W. Hummel and M.-R. Kula, Recent developments in separation and purification of biomolecules, Ann. N.Y. Acad. Sci. 413:270 (1983)PubMedCrossRefGoogle Scholar
  2. 2.
    M.-R. Kula, K.H. Kroner and H. Hustedt, Purification of enzymes by liquid-liquid extraction, Advan. Biochem. Eng. 24:73 (1982)Google Scholar
  3. 3.
    J. Rudge, M.A. Desai, S.A. Shojaosadaty and A. Lyddiatt, Continuous culture of murine hybridomas with integrated recovery of monoclonal antibodies, in: “Modern Approaches to Animal Cell Technology,” R.E. Spier and J.B. Griffiths, eds., Butterworth, LondonGoogle Scholar
  4. 4.
    U.K. Laemmli, Cleavage of structural protein during the assembly of the head of bacteriophage T4, Nature, 227:680 (1970)PubMedCrossRefGoogle Scholar
  5. 5.
    M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding, Anal. Biochem. 72:249 (1976)CrossRefGoogle Scholar
  6. 6.
    S.A. Shojaosadaty, Application of affinity HPLC to the monitoring and quantitation of biological process operations, Conference Proceedings EFB4. Amsterdam 3:136 (1987)Google Scholar
  7. 7.
    D.T. Plummer, Introduction to practical biochemistry (2nd edition), McGraw-Hill, London (1978)Google Scholar
  8. 8.
    D. Herbert, P.J. Phipps and R.E. Strange, in: “Methods in Microbiology”, J.R. Norris and D.W. Ribbons, eds., Vol. 5B:285, Academic Press, London (1971)Google Scholar
  9. 9.
    B. Kassell and P.A. Meitner, Bovine pepsinogen and pepsin, in: “Methods in Enzymol.,” Academic Press, London (1970)Google Scholar
  10. 10.
    B. Skoog, Determination of poly(ethylene glycols) 4000 and 6000 in plasma preparations, Vox. Sng. 37:345 (1979)CrossRefGoogle Scholar
  11. 11.
    M.A. Desai, A. Lyddiatt, J. Rudge, A.W. Sansome-Smith and A.B. Stevens, Biochemical and physical characterisation of a composite solid phase developed for large scale biochemical adsorption, Conference Proceedings EFB4 Amsterdam 2:517 (1987)Google Scholar
  12. 12.
    K.H. Kroner, H. Hustedt and M.-R. Kula, Extractive enzyme recovery-economic considerations, Process Biochem. 19:67 (1984)Google Scholar
  13. 13.
    C.J.O.R. Morris and P. Morris, Countercurrent distribution, in: “Separation Methods in Biochemistry,” Pitman, London (1976)Google Scholar
  14. 14.
    P.O. Hedman, Protein adsorbents intended for use in aqueous two-phase systems, Anal. Biochem. 138:411 (1984)PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Jon G. Huddleston
    • 1
  • Andrew Lyddiatt
    • 1
  1. 1.Biochemical Recovery Laboratory, Department of Chemical EngineeringUniversity of BirminghamBirminghamUK

Personalised recommendations