muramidase and divinyl-benzene

A strong cation-exchange separation material has been prepared from monodisperse divinylbenzene particles modified by a "grafting to" approach, utilizing as anchoring points epoxy groups introduced onto the surface of the particles via oxidation of residual vinyl groups. The grafted chains consisted of thiol-terminated telomers of sulfopropyl methacrylate prepared by iniferter mediated polymerization, and grafting was performed by reaction of the corresponding thiolate anion with the surface epoxy groups. Attachment through epoxy moieties that were subsequently converted into 2,3-propanediol groups increased the hydrophilicity of the polymeric particles and incubation experiments showed no signs of the proteins denaturing on the column during an extended contact time of 1 h at room temperature. The performance of the grafted material was demonstrated by the chromatographic separation of cytochrome C, lysozyme, myoglobin, and ribonuclease A, in a cation-exchange mode.

Topics: Chromatography, Ion Exchange; Cytochromes c; Ion Exchange Resins; Methacrylates; Microscopy, Electron, Scanning; Muramidase; Myoglobin; Particle Size; Polymers; Proteins; Ribonuclease, Pancreatic; Sulfhydryl Compounds; Surface Properties; Vinyl Compounds

A poly(styrene-divinylbenzene) chromatography matrix, CG1000sd (TosoHaas) has been modified by the adsorption and crosslinking of poly(vinyl alcohol) (PVA) to create a matrix suitable for the attachment of dye ligands for the adsorption of lysozyme. However, it is shown that there was limited recovery and repeated drops in capacity with adsorption of human serum albumin (HSA). The effect of changing the nature of the PVA crosslinking on the HSA binding characteristics was studied, as well as the effect of using differing dye ligands. The total amount of irreversible HSA binding decreased with greater crosslinking and there were large differences in HSA adsorption characteristics between differing dye types.

Topics: Adsorption; Chemical Phenomena; Chemistry; Chromatography, Liquid; Coloring Agents; Cross-Linking Reagents; Glutaral; Ligands; Muramidase; Polystyrenes; Polyvinyl Alcohol; Proteins; Spectrophotometry, Ultraviolet; Vinyl Compounds

Poly(styrene-divinylbenzene) (PS-DVB) chromatography matrices, CG1000sd 20-50 microns (TosoHaas), PLRP4000s 15-25 microns, PLRP4000s 50-70 microns (Polymer Laboratories) have been modified by the adsorption and crosslinking of poly(vinyl alcohol) (PVA) to create a matrix suitable for the attachment of dye ligands. The adsorption capacities of lysozyme and HSA on these Procion Yellow HE-3G dyed PVA modified PS-DVB matrices were measured at various flow-rates and the capacities were compared with a Procion Yellow HE-3G dyed OH-activated POROS 20, 20-micron matrix (PerSeptive Biosystems). The adsorption of small proteins was not hindered by the smaller pores of the CG1000sd beads, but as protein size increased, and at high flow-rates, a high mass transfer rate became more dependent on large pore size and small particle diameter.

Topics: Adsorption; Chromatography; Cross-Linking Reagents; Fluorescent Dyes; Kinetics; Muramidase; Particle Size; Polystyrenes; Polyvinyl Alcohol; Porosity; Proteins; Spectrophotometry, Ultraviolet; Triazines; Vinyl Compounds