Affinity dextran gels

Recovery and Purification. The dalbaheptides are present in both the fermentation broth and the mycelial mass, from which they can be extracted with acetone or methanol, or by raising the pH of the harvested material, eg, to a pH of 10.5—11 for A47934 (16) (44) and A41030 (41) and actaplanin (Table 2) (28). A detailed review on the isolation of dalbaheptides has been written (14). Recovery from aqueous solution is made by ion pair (avoparcin) or butanol (teicoplanin) extraction. The described isolation schemes use ion-exchange matrices such as Dowex and Amberlite IR, acidic alumina, cross-linked polymeric adsorbents such as Diaion HP and Amberlite XAD, cation-exchange dextran gel (Sephadex), and polyamides in various sequences. Reverse-phase hplc, ion-exchange, or affinity resins may be used for further purification (14,89). 

Cibacron Blue is a blue, polyaromatic, sulfonated dye (Fig. 6). It can be attached, as an affinity li nd, to solid matrix supjmrts (e.g. dextran, agarose) by the reaction of the triazine ring with free hydroxyl groups of the supports. The conditions of this triazine coupling method have been described by Bohme Such a dye affinity sorbent is also produced commercially, e.g. under trade name Blue Sepharose CL-6B (Cibacron Blue F3G-A covalently bound to the cross-linked agarose gel Sepharose CL-6B ) by Pharmacia, Sweden. 

Classical LLPC using aqueous-aqueous polymer systems based on Albertsson s [9] PEG-dextran system has provided a versatile tool for the separation of proteins and nucleic acids, thus increasing the arsenal of biopolymer purification methods currently dominated by gel filtration, ion-exchange chromatography, and affinity chromatography RPC. The technique operates... 

The main drawback to the widespread use of polymer-polymer aqueous two-phase extraction has been the high cost of fractionated dextran. Crude dextran has been used with some success for the purification of enzymes but is much too viscous for many applications. Conversely, polymer-salt systems have relatively low viscosities, separate rapidly, and are inexpensive. Unfortunately, they lack selectivity and cannot be used for affinity partitioning in most cases since the high salt concentrations interfere with the protein-ligand interaction. The starch derivatives are reasonable alternatives for bottom phase polymers but have been hampered by low solubilities and the tendency for gel formation. Tjemeld has reported that chemically modified starches i.e. hydroxypropyl starch... 

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