Sepharose fractionation ranges

TABLE 16.6 Producers (Pharmacia Biotech) Specification of Fractionation Ranges of 2,3-Dibromopropanol Cross-Linked Agarose Derivative-Based Sepharose CL Gels... 

TABLE 2. Range of Sephadex and Sepharose Gels Manufactured by Pharmacia Showing Their Fractionation Range and Exclusion Limit... 

Separate lipid-encapsulated ones from free drug molecules using an appropriate column. In-house, for separation of free ohgonucleotides from encapsulated ones, a Sepharose CL-2B, fractionation range 70,000-40,000,000, is used. 

Combining Sepharose CL 2B and CL 4B singnificantly increases resolution in the low dp range of broad distributed samples. As an example, wild-type potato starch and two fractions of this sample differing in their branching... 

Purified C5-1 has been obtained from alfalfa leaf extracts by affinity chromatography on either a human IgG-Sepharose column or a Streamline rProtein A-Sepharose column. Interestingly, the purified product obtained with these two methods differed significantly. As shown in Fig. 1.5 a, the antibody fraction obtained from the human IgG column contained a mixture of different intermediate assembly forms of the heavy (H) and light (L) chains, ranging from H2 to the fully assembled H2L2 form. 

The B875 kinase activity present in the supernatant fraction was then isolated by affinity chromatography on a Sepharose-ATP column and gel filtration. The purified enzyme was present in only low amounts but rapidly phosphorylated intact chromatophores in the presence of DBMIB or isolated B875 light harvesting complexes, as well as histone V-S (Fig. 5). Its molecular weight was in the range of 18 kD. 

By ion-exchange chromatography on DEAE-Sepharose FF, the two ammonium sulfate fractions (primary fractions) were separated into subfractions (Fig. 2.8). Seven subfractions showed protective activity in E. coli inactivated by UV light. When the subffactions were analyzed by SDS polyacrylamide gel electrophoresis and HPLC, it was found that the subfraction AF2-2.5 contained mostly a single protein, whereas the other subfractions were composed of mixtures of proteins and peptides of different molecular weights. The molecular weight of subfraction AF2-2.5 was determined at 44 2 kDa (Zinchenko et al, 1998). The protective activity of subfraction AF2-2.5 in UV-inactivated E. coli was dependent on its concentration in the range of 10-60 pg of protein per ml. 

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