Liquid-Liquid Partition Chromatography (LLPC)

  • Ulla-Britt Hansson
  • Christer Wingren
Part of the Methods in Biotechnology™ book series (MIBT, volume 11)


Liquid-liquid partition chromatography (LLPC) is a column chromato-graphic approach of aqueous two-phase partitioning that offers new and unique possibilites to solve separation and fractionation problems (1, 2, 3, for review,) see ref. 4). The bottom phase of the two-phase system is adsorbed onto a support and packed into a column, which is eluted with the corresponding top phase Fig. 1). The LLPC method is a rapid (≤ 2 h), sensitive (0.1-100 µg protein is required) analytical method with which even large, flexible, and heterogeneous biomolecules can be analyzed in aqueous solution. The obtained results are highly reproducible (the relative standard deviation of K is <- 5%) and conventional chromatographic equipment available in most laboratories can be used Fig. 1).
Fig. 1.

Schematic illustration of the LLPC setup. A typical chromatogram for the reference proteins, peroxidase and myoglobin, obtained on LiParGel 650 (300×8 mm I.D.) in a 4.4% (w/w) PEG 8000/6.2% (w/w) dextran T 500,50 mM sodium phosphate, 0.1 M NaCl, 0.1 M glycine, pH 7.0, at a flow rate of 0.12 mL/min (20°C) is shown.


Retention Volume Reference Protein Bottom Phase Buffer Composition Ideal Partitioning 
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Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Ulla-Britt Hansson
    • 1
  • Christer Wingren
    • 2
  1. 1.Department of Biochemistry, Center for Chemistry and Chemical EngineeringLund UniversityLundSweden
  2. 2.Department of BiochemistryCenter for Chemistry and Chemical Engineering, Lund UniversityLundSweden

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