Continuous Affinity Partitioning in a Poly(Ethylene Glycol)-Reppal Pes Aqueous Two-Phase System
The starch-based polymer, Reppal Pes, has been used to form two-phase affinity systems with dyed-poly(ethylene glycol). Lactate dehydrogenase has been purified from a crude rabbit muscle extract in these systems using the triazine dyes Cibacron blue F3G-A and Procion Orange H-ER bound to poly(ethylene glycol). Processing parameters have been optimised in small-scale batch experiments and the results used to formulate systems for continuous affinity partitioning of rabbit muscle lactate dehydrogenase in a simple two-stage mixer-settler unit. Polymer-bound ligand from the eluting stage has been recycled into the affinity partitioning stage, thereby reducing the amount of immobilised ligand required.
KeywordsProduct Stream Bottom Phase Settling Chamber Pyruvate Kinase Activity Purification Ratio
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
A. Cordes and M.-R. Kula, Process design for large-scale purification of formate dehydrogenase from Candida boidinii by affinity partition, J. Chromatography
376:375 (1986)CrossRefGoogle Scholar
A. Veide, A.-L. Smeds and S.-O. Enfors, A process for large-scale isolation of ß-galactosidase from E. coli in an aqueous two-phase system, Biotechnol. Bioeng.
25:1789 (1983)PubMedCrossRefGoogle Scholar
C.R. Lowe, New developments in downstream processing, J. Biotechnol.
1:3 (1984)CrossRefGoogle Scholar
P.-F. Fauquex, H. Hustedt and M.-R. Kula, Phase equilibration in agitated vessels during extractive enzyme recovery, J. Chem. Tech. Biotechnol.
35B:51 (1985)Google Scholar
G. Johansson, Aqueous two-phase systems in protein purification, J. Biotechnol.
3:11 (1985)CrossRefGoogle Scholar
K.H. Kroner, H. Hustedt and M.-R. Kula, Evaluation of crude dextran as phase-forming polymer for the extraction of enzymes in aqueous two-phase systems in large-scale, Biotech. Bioeng.
24:1015 (1982)CrossRefGoogle Scholar
F. Tjerneld, S. Berner, A. Cajarville and G. Johansson, New aqueous two-phase system based on hydroxypropyl starch useful in enzyme purification, Enzyme Microb. Technol.
8:417 (1986)CrossRefGoogle Scholar
R. Kaul and B. Mattiasson, Extractive bioconversion in aqueous two-phase systems. Production of prednisolone from hydrocortisone using Arthrobacter simplex as catalyst, Appl. Microbiol. Biotechnol.
24:259 (1986)CrossRefGoogle Scholar
B. Mattiasson and T.G.I. Ling, Efforts to integrate affinity interactions with conventional separation technologies. Affinity partition using biospecific Chromatographic particles in aqueous two-phase systems, J. Chromatography
376:235 (1986)CrossRefGoogle Scholar
K.H. Kroner, H. Hustedt, S. Granda and M.-R. Kula, Technical aspects of separation using aqueous two-phase systems in enzyme isolation processes, Biotech. Bioeng.
20:1967 (1978)CrossRefGoogle Scholar
M. Joelsson and G. Johansson, Sequential liquid-liquid extraction of some enzymes from porcine muscle using polymer-bound triazine dyes, Enzyme Microb. Technol.
9:233 (1987)CrossRefGoogle Scholar
G. Johansson and M. Joelsson, Preparation of Cibacron Blue F3G-A poly(ethylene glycol) in large-scale for use in affinity partitioning, Biotech. Bioeng.
27:621 (1985)CrossRefGoogle Scholar
L.A. Decker, ed., “Worthington Enzyme Manual,” Worthington Biochemical Corp., Freehold NJ (1977)Google Scholar
“Biochemica Information I,” Boehringer Mannheim GmbH, Mannheim (1973)Google Scholar
M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding, Anal. Biochem.
72:248 (1976)PubMedCrossRefGoogle Scholar
G. Johansson and M. Joelsson, Liquid-liquid extraction of lactate dehydrogenase from muscle using polymer-bound triazine dyes, App. Biochem. Biotech.
13:15 (1986)CrossRefGoogle Scholar
S.D. Flanagan and S.H. Barondes, Affinity partitioning — a method for purification of proteins using specific polymer-ligands in aqueous polymer two-phase systems, J. Biol. Chem.
250:1484 (1975)PubMedGoogle Scholar
C.G. Hill, Jr., “An Introduction to Chemical Engineering Kinetics and Reactor Design,” Wiley, New York (1977)Google Scholar
© Plenum Press, New York 1989