Bead cellulose derivatives as supports for immobilization and chromatographic purification of proteins.
Characteristic data are presented for Divicell, a macroporous bead cellulose with excellent flow parameters. The preparation of Divicell derivatives and their properties are described with respect to their application as chromatographic supports. The ion exchangers Divicell DEAE and Divicell CM were manufactured in two types with different exclusion limits and an available capacity for proteins of up to 100 mg/ml gel. Divicell Blue is a bead cellulose with covalently bound Cibacron Blue F3G-A and was found to be a very suitable adsorbent for the selective separation and purification of human serum albumin. Activation of Divicell with sodium periodate, epichlorohydrin and 5-norbornene-2,3-dicarboximido carbonochloridate provided activated supports used for immobilization of ligands in organic solvents and in aqueous solutions. Coupling of amines, diamines, amino acids, carbohydrates and proteins is described. The immobilized ligands retained their biological activity as determined by their specific adsorption of proteins. Divicell alkyl derivatives were tested in hydrophobic interaction chromatography with bovine serum albumin as a model. Examples are presented of the application of Divicell derivatives to the purification of biomacromolecules such as immunoglobulins and lectins by affinity chromatography. The results were comparable to those obtained using the corresponding Sepharose-derived absorbents.
Central Institute of Molecular Biology, Berlin, Germany.
H F
HF
Boeden
K
K
Pommerening
M
M
Becker
C
C
Rupprich
M
M
Holtzhauer
F
F
Loth
R
R
Müller
D
D
Bertram
D000818:Animals; D002482:Cellulose; D002645:Chickens; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D008854:Microscopy, Electron; D010047:Ovalbumin / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D011817:Rabbits; D012709:Serum Albumin / Q000302:isolation & purification; D014909:Wheat Germ Agglutinins / Q000302:isolation & purification
D007074:Immunoglobulin G; D011506:Proteins; D012709:Serum Albumin; D014909:Wheat Germ Agglutinins; D002482:Cellulose; D010047:Ovalbumin