Magnetic aqueous two-phase separation in preparative applications.
Magnetic aqueous two-phase separation is a new technique to speed up the separation of aqueous two-phase systems (Anal. Biochem. 1987, 167, 331-339). It is based on the addition of magnetically susceptible material (e.g. 1-micron iron oxide particles) which induces rapid phase separation when a mixed system is placed in a magnetic field. The technique has been applied to a number of two-phase systems. The time for phase separation was decreased by a factor of 5-240,000, with the largest improvement for systems containing high concentrations of protein and for systems with viscous or nearly isopycnic phases. An apparatus for preparative multistage extraction with magnetic separation was constructed and tested on glycolytic enzymes present in a yeast extract using a dextran/Cibacron blue-polyethylene glycol system. The presence of iron oxide particles did not adversely affect the extracted enzymes. An electromagnet-based apparatus for continuous phase separation on a larger scale was also designed. A phase system containing crude dextran and unpurified cell homogenate was effectively processed. The apparatus also allowed effective separation when the phase containing iron oxide particles was only a small fraction (4%) of the total phase system.
Flygare
S
S
Department of Pure and Applied Biochemistry, University of Lund, Sweden.
Wikström
P
P
Johansson
G
G
Larsson
P O
PO
D000426:Alcohol Dehydrogenase / Q000302:isolation & purification*; D005290:Ferric Compounds; D005656:Fungal Proteins / Q000302:isolation & purification; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification*; D006593:Hexokinase / Q000302:isolation & purification*; D008280:Magnetics; D010732:Phosphofructokinase-1 / Q000302:isolation & purification*; D011108:Polymers; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D005290:Ferric Compounds; D005656:Fungal Proteins; D011108:Polymers; C000499:ferric oxide; D000426:Alcohol Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D010732:Phosphofructokinase-1