Thin-layer chromatography of malto-oligosaccharides.
Abstract Procedures for the separation, location and elution of malto-oligosaccharides are described. Best separations were obtained on glass plates (20 × 46 × 0.5 cm) coated with Kieselguhr G, serially developed in solvents of varying proportions of x-butanol, pyridine and water. Major zones were temporarily visualized by exposing the plates to iodine vapors. The located zones were eluted from the thin-layer plates into fiberglass filter pieces with water. Radioactivity of the zone can be determined directly by placing the filter in a toluene-based scintillation fluid and counting in a scintillation counter. After radioactivity determination, the carbohydrates can be eluted from the filter with water and quantitatively measured.
Relationship between phosphate content and immunochemical properties of subfractions of bakers' yeast mannan
The mannan of bakers' yeast (Saccharomyces cerevisiae) was fractionated on a column of diethylaminoethyl-Sephadex into five subfractions. Phosphate content of these mannan subfractions was proportional to the concentration of NaCl solutions used in the chromatographic separation. Quantitative precipitin reactions showed that the serological reactivities of the subfractions were proportional to the content of phosphate. The result of acetolysis study showed that the amounts of mannotetraose and phosphate-containing oligosaccharide fractions increased proportionally to the acidity, whereas the amount of mannose decreased inversely. The results from quantitative precipitin reaction tests and acetolysis study demonstrated that both phosphate contents and multiplicity of branching moieties of mannan subfractions increased proportionally, i.e., micro-heterogeneity concerning the acidity comprised in the parent bulk mannan is not attributable merely to the coexistence of molecular species containing different amounts of phosphate but also to the presence of more of the branching moieties.
Adsorption of immunoglobulin Y in supermacroporous continuous cryogel with immobilized Cu(2+) ions.
The influence of temperature, pH and ionic strength on the adsorption of Immunoglobulin Y (IgY) in IDA-Cu(2+) cryogel system was studied by batch equilibrium measurements. The adsorptive equilibrium data were obtained at 17 and 27°C, pH 5.0 and 6.5, and at ionic strength of 50 and 200mmolL(-1) NaCl. Langmuir, Freundlich and Langmuir-Freundlich models were fitted to equilibrium data, while the enthalpy of adsorption of IgY in IDA-Cu(2+) cryogel system was calculated through Van't Hoff analysis. The binding of IgY on cryogel was stronger at 27°C and lowest pH and ionic strength values, with apparent maximum adsorption capacity of 27mgg(-1). The adsorption of protein in the resin was spontaneous in all analyzed cases. The results provide valuable information to enable the improvement of IgY purification processes.