Coated silica supports for high-performance affinity chromatography of proteins.
Polymer-coated silica supports are potentially good stationary phases for high-performance affinity chromatographic separations of proteins. Silica beads have been coated with a polysaccharide (dextran or agarose), substituted by a calculated amount of positively charged diethylaminoethyl functions in order to neutralize the negatively charged silanol groups of silica and to facilitate the formation of a hydrophilic polymeric layer on the inorganic surface. The silica-based supports were prepared in two steps. First, the silica was impregnated with a solution of diethylaminoethylated polymer, and then the coating polymer was crosslinked in order to avoid leakage of the polymeric layer. The supports present minimal non-specific interactions with proteins, as tested by high-performance size-exclusion chromatography. These coated silica supports were coupled with active ligands, such as protein A, concanavalin A and heparin, by conventional coupling methods. The resulting affinity stationary phases were tested by the elution of proteins in order to study their performance in high-performance affinity chromatography.
Zhou
F L
FL
L.R.M. CNRS UA502, University Paris-Nord, Villetaneuse, France.
Muller
D
D
Santarelli
X
X
Jozefonvicz
J
J
D000990:Antithrombin III / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods; D006493:Heparin / Q000032:analysis; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D008024:Ligands; D010047:Ovalbumin / Q000032:analysis; D011134:Polysaccharides / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D013917:Thrombin / Q000032:analysis
D007074:Immunoglobulin G; D008024:Ligands; D011134:Polysaccharides; D011506:Proteins; D000990:Antithrombin III; D006493:Heparin; D010047:Ovalbumin; D013917:Thrombin