[Synthesis and evaluation of high performance gel filtration chromatography packing of KH-s-GFC300].
A novel packing for high performance gel filtration chromatography (GFC) was synthesized and characterized. High porosity silica prepared by base-dissolving method was used as the matrix. gamma-(2,3-Epoxy propoxyl) propyltrimethoxysilane was used as the ligand and covalently bonded onto the silica matrix. After acidic hydrolysis, the epoxy groups were converted to the diol groups. Because a condensation tube filled with water at 70 degrees C was used in the grafting reaction, the resulting methanol could easily be discharged from the reaction system to shift the reaction equilibrium to achieve high ligand density. The hydrolysis condition greatly affects ligand density and column efficiency. The high column efficiency is observed when the ligand density is between 2.6 and 3.5 +/- mol/m2. Several proteins, such as cytochrome C, chymotrypsin, ovalbumin, bovine serum albumin, aldolase, ferritin, insulin, gamma-globulin, phosphorylase, actin, carbonic anhydrase, were used to characterize the separation properties of the resulting high performance GFC column. It was shown that the excluded limit of relative molecular mass for the separation of bio-molecules was 300 000. The recovery yield of bovine serum albumin was 99%.
Center for Molecular Science, Institute of Chemistry, The Chinese Academy of Sciences, Beijing 100080, China. zhaorui@iccas.ac.cn
Rui
R
Zhao
Xinhui
X
Li
Guoquan
G
Liu
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel / Q000295:instrumentation*; D011108:Polymers / Q000138:chemical synthesis* / Q000737:chemistry*; D016062:Porosity; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D012822:Silicon Dioxide / Q000737:chemistry
D011108:Polymers; D011506:Proteins; D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide