Separation of enzymes

Basic technology for membrane separation of biomolecules was invented in the United States, but the West Germans and the Japanese lead in its application to separations of enzymes and amino acids from complex mixtures. Japanese... 

Meindersma and Kuczynski (1996) compared conventional filtration with a filter aid and membrane technology for the separation of enzymes. In cases in which the enzyme is sufficiently cheap for once-through use filtration is the process of choice, whereas for... 

Liquid Formulations. The successful application of enzymes in liquid formulations presents several technical obstacles that are not encountered in powders and granules. These problems stem from the fact that liquid products are complex aqueous solutions, and physical separation of enzymes from other potentially inactivating ingredients is impractical. Necessary ingredients for a viable end-product can affect the physical and chemical stability of enzymes. In addition, as other types of enzymes are added to formulations that already contain proteases, proteolytic degradation of enzymes is a concern. 

Peroxidase, in combination with phenolic compounds, utilizes hydrogen peroxide to bring about oxidation. The enzymes do not act in intact fruits because of the physical separation of enzyme and substrate. Mechanical damage, rot, or senescence lead to cellular disorganization and initiate decomposition. Inhibition of the enzymes in vegetables is achieved by blanching with steam or by... 

Inada et al. succeeded in combining enzyme on synthesized magnetic particles. Thermosensitive magnetic particles that are useful for separation of enzyme and biomaterials have been reported. Furthermore, since magnetic polymeric microsphere particles were developed for labeling and separation of biocells, " studies into the applications of magnetic particles in the biology and medical fields are under way. ... 

Bermex, L.G. Freitag, R. Affinity-based Interactions on Disks for Fast Analysis, Isolation and Conversion of Biomolecules. In Methods for Affinity-Based Separations of Enzymes and Proteins Gupta, M.N., Ed. Birkhauser Basel, 2002 Chapter 7. 

The separation of proteins and enzymes is performed with ultrafiltration membranes. Branger et al. [93] use Carbosep Ml and M4 (40,000 and 20,000 Dalton respectively) for the separation of enzyme hydrolysates. The fluxes with these membranes compare favourably with polymeric membranes 37-102 l/m hvs. 7-41 l/m h. 

Cortex Biochem has found interesting applications for magnetizahle particles in analytical fields. Particles of the analytic aid are prepared Ifom a combination of magnetizable materials (iron oxide) and absorbing material (e.g., charcoal, polyacrolein, ion exchange, cellulose). The particles are dispersed in a biological sample to selectively absorb required compounds. After absorption was accomplished, particles with absorbed substance are removed from solution by a magnetized rod. The materials are used for separation of enzymes, protein, cells or bacteria. 

Ion-exchange HPLC separation of enzymes has been a particularly active area of research, mainly because of the clinical usefulness of isoenzyme measurements. Most separations employ a conventional colorimetric or fluorometric assay on the eluent (post-column) to detect resolved isoenzymes (F9). 

Ohlson, S., Hansson, L., Larsson, P-O., and Mosbach, K., High performance liquid affinity chromatography (HPLAC) and its application to the separation of enzymes and antigens. FEBS Lett. 93, 5-9 (1978). 

The method has been used for the separation of enzymes, hormones, RNA-DNA hybrids, ribosomal subunits, subcellular organelles, for the analysis of distribution of samples of polysomes and for lipoprotein fractionation. 

This packing has high capacity and has been found useful for the separation of enzymes and other proteins. The ion exchangers suitable for the separation of biopolymers are summarized in the lower part of Table 4.5.2. 

Unlike mass spectroscopy, gel electrophoresis does not provide a quantitative value for the amount of given protein. However, it provides a low cost and relatively rapid method for the analysis of multiple proteins in a specimen, especially when implemented as a capillary electrophoresis system. Therefore, it has been used for the separation of enzymes (e.g., creatinine phosphokinase), mucopolysaccharides, plasma, serum, cerebrospinal fluid, urine, and other bodily fluids [13]. It is also used for quality control applications for the manufacturing of biological compounds to verify the purity or to examine the manufacturing yield [14]. 

Sharma, A., Mondal, K., and Gupta, M. N. 2003. Separation of enzymes by sequential macroaffinity ligand-facilitated three-phase partitioning. J. Chromatogr. A 995 127-134. 

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