USE OF A TEMPERATURE-INDUCED PHASE-FORMING DETERGENT (TRITON-X-114) AS LIGAND CARRIER FOR AFFINITY PARTITIONING IN AN AQUEOUS 3-PHASE SYSTEM

The use of a non-ionic detergent (Triton X-114) as ligand carrier in a queous two-phase extraction of proteins was studied. Triton X-114 was modified with Cibacron Blue and partitioned in a poly(ethylene glycol) -hydroxypropyl-starch system. When an excess of detergent was used, a three-phase system with a detergent-rich middle phase was formed. The detergent-dye conjugate partitioned to this detergent-rich phase. This detergent-dye conjugate was used as an affinity ligand for lactate de hydrogenase. The enzyme was recovered by harvesting the detergent-rich -conjugate-containing phase and subjecting it to temperature-induced p hase separation, whereby the detergent and conjugate formed a separate phase, leaving the enzyme in the water-rich phase.