Two-Phase Affinity Partitioning of Animal Cells: Implications of Multipoint Interactions

  • Ashok Kumar
  • Masamichi Kamihira
  • Bo Mattiasson
Part of the Methods and Tools in Biosciences and Medicine book series (MTBM)

Abstract

Partitioning of biological macromolecules, e. g., proteins, cells, organelles and membranes, has become an established method for their separation and fractionation [1]. The success of partitioning depends on numerous intrinsic properties such as size, electrochemical properties, hydrophobic and hydrophilic surface properties, conformation properties, and other factors. Extrinsic parameters influencing partitioning of macromolecules are: type and molecular weight of the polymers used, concentration of polymers, type of buffer components, ionic strength, pH of the buffer, temperature and others. A selective partitioning of a target biomolecule to a specific phase can be achieved by introducing an affinity ligand to the system (ATPS) [2] Affinity partitioning combines the capability of biological macromolecules to partition in aqueous two-phase systems with the principle of biorecognition. The affinity ligands are introduced into the systems which are coupled either to phase-forming polymers or ligand carriers which can be partitioned completely to one of the phases formed. Systems are described where specific protein molecules are enriched in one phase with high yield and also some times in high purity. The phenomena behind such a separation are very much the same if, instead of soluble proteins, membrane-bound proteins, or even organelles or cells are partitioned. One of the very attractive features of partitioning in ATPS is that it does not make much of a difference if the target is a soluble protein or a particulate structure. Because of this fact we describe here the separation of cells, but it could equally well have been other subcellular structures or soluble molecules. The difference is found in the possibility for multipoints of attachment.

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References

Further Reading

  1. Hatti-Kaul R (ed) (2000) Aqueous two-phase systems. methods and protocols. Humana Press, Totowa, NJ, USAGoogle Scholar
  2. Albertsson P-Å (ed) (1986) Partition of cell particles and macromolecules,3rd ed., Wiley (Interscience), New YorkGoogle Scholar
  3. Walter H, Brooks D E, Fisher D (eds) (1985) Partitioning in aqueous two-phase systems. Theory, methods, uses and applications to biotechnology. Academic Press, Orlando, FLGoogle Scholar
  4. Walter H, Johansson G (eds) (1995) Methods in enzymology: aqueous two-phase systems, vol. 228. Academic Press, San Diego, CA.Google Scholar

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Copyright information

© Springer Basel AG 2002

Authors and Affiliations

  • Ashok Kumar
  • Masamichi Kamihira
  • Bo Mattiasson

There are no affiliations available

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