Liquid-Liquid Partition Chromatography (LLPC)
互联网
493
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 – 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 �g 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.
