Active extract purification

A method for extraction, purification and preconcentration of dialkyldimethylammo-nium compounds and other detergents before determining their concentration in sewage water and activated sludge was described. It consists of a series of LLE and LC operations, the details of which are dependent of the original matrix, and end analysis was by HPLC-ELCD406. 

Additional assays included a selectivity evaluation and cytotoxicity assay where both good selectivity and low cytotoxicity were documented. The extract was also tested on unrelated viruses and other influenza viruses. Extract A4 was the only one to maintain the activity through secondary screening, and with that as a target, purification of the active extract commenced. 

However, we have observed that values obtained with crude extracts were only qualitative. Often, they did not accurately estimate the quantities of the individual enzymes present. Inhibitors were typically present that caused the underestimation of certain enzymes ie.g., ligninases Table II) and that could potentially mask less dominant enzymes. Also, certain polysaccharidases e.g., hemicellulases) were often overestimated due to the action of non-specific or synergistic enzymes e.g., other hemicellulases or cellulases) (9,14), This artifact resulted in low apparent recovery of a given activity and only moderate increases in specific activity upon purification of the major corresponding enzyme present, in spite of the fact that SDS polyacrylamide gels indicated good recovery and substantial removal of contaminants (14),... 

Intact antibodies with biologically active glycosylation profiles, crucial for the effector functions, require eukaryotic expression (in vitro or in vivo). These circumstances have inspired many scientists to find effective methods for production, as well as methods for the selection of the best extraction-purification methods. 

Antibodies are very diverse in molecular properties, chemical characteristics, and biological activity. Purification strategies are therefore also diverse, since they are based on a large variety of molecular interactions. Antibodies have several common properties that are frequently exploited from the initial feedstock. Knowledge about the nature and concentration of impurities is the key to success. Therefore, the initial composition of feedstock is important when designing the separation process. The expression system can be selected to simplify the extraction-purification procedures. 

When the ultimate objective is to produce bioactive peptides for particular purposes, such as antioxidative or antihypertensive activities, the purification of target peptides from protein hydrolysate can be carried out using UF membranes with or without chromatographic techniques. Jun et al (2004) reported successful preparation of protein hydrolysates from yellowfin sole frame by first using extracted mackerel intestine crude enzyme at pH 10.0 and 50 °C, followed by treatment with pepsin at pH 2.0 and 37 °C. The resultant hydrolysate was further fractionated through five different UF membranes with... 

The assay has been used with mitochondrial extracts prepared from freeze-thawed mitochondria from rabbit and pigeon, and to monitor enzyme activity during purification. 

In environmental chemical analytical procedures such as extraction, purification, and fractionation with basic aluminum oxide and activated carbon,... 

Purffication. Lucidol material contains 76-90% of active reagent. Purification has been accomplished by fractionation - and by refluxing in an azeotrope separator until homogeneous and then distillation. Kochi and Mocadlo vacuum distilled Lucidol material several times and obtained a product of greater than 99% purity. Bartlett and McBride extracted Borden material with cold 15% aqueous potassium hydroxide, followed by neutralization with solid ammonium chloride and vacuum distillation (98.4% pure). 

Washed cell pastes were diluted with fresh buffer using a ratio of 3 ml of buffer for each 1 g of cell paste. A French Cell Press operated at 16,000 lb/in was used to disrupt cells. The crude enzyme preparation was then centrifuged at 10,444 X g for 30 min at 0°C, and the supernatant was retained as a cell-free extract (CFE). This extract was carried through a series of activation and purification steps and the methlonlnase activity was assayed after each step using a modification of the method of Tanaka t al. (52). Protein was assayed by the dye-binding method of Bradford (62), and enzyme solutions were dialyzed in tubing prepared by procedures described by Brewers at al. (63). 

A crucial point when discussing economics of phase systems is the loading of the system. The more biological material that can be loaded, lesser is the cost of phase system per unit processed. Furthermore, extensive studies have been carried out concerning recirculation of the phase components. The latter efforts have mainly been focused on PEG/salt systems, and also on the polymer/polymer phase systems employing affinity ligands. Kula and coworkers reported the reuse of the salt phase when carrying out extractive purification (37). They showed that the phases could be recycled 4 times without any marked loss in performance of the phase separation and in specific activity of the purified protein. 

Dtdemnins, Biologically active depsipeptides extracted from a Caribbean tunicate (sea squirt) family Didem-nidae. genus Trididemnum. The three components, didem-nins A, B and C are weakly basic compds with antiviral, antitumor activity. Didemnin A is the most abundant, whereas didemnin B is generally the most active. Extraction and purification K., L. Rinehart. Jr., Eur. pat. Appl. 48,149 and U.S. pat. 4,493,796 (1982, 1985 both to Univ. Illinois). Structure determn K, L. Rinehart. Jr. et ah, J. Am. Chem. Soc. 103, 1857 (1981). Revised structure and total synthesis K. L. Rinehart, Jr- et ah. ibid. 109, 6846 (1987). See also U. Sebmidt et ah. Tetrahedron Letters 29, 3057 (1988) eidem, ibid. 4407. Crystal structure of B M. B. Hossain el ah. Proc. Nat. Acad. Sci. USA 85, 4118 (1988). Efficient total synthesis of A and B Y. Hamada et ah. J. 

You Q,Yin X, Zhang S, Jiang Z. Extraction, purification, and antioxidant activities of polysaccharides from Tricholoma mongolicum Imai. Carbohydr Polym 2014 99 1-10. 

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