High-performance liquid chromatography of amino acids, peptides and proteins. XCII Thermodynamic and kinetic investigations on rigid and soft affinity gels with varying particle and pore sizes.
In these investigations Cibacron Blue F3GA was immobilized on soft gels, porous silicas, and non-porous glass beads. Hen egg white lysozyme, human serum albumin and yeast alcohol dehydrogenase were used as adsorbates with the dye-affinity sorbents. Batch experiments with continuous monitoring of protein concentration were employed to evaluate thermodynamic and kinetic behaviour of these proteins in finite bath systems. The observed adsorption kinetic rates of interaction of the above proteins with each of the dye-affinity sorbents were found to decrease with increasing protein molecular weight. Equilibration times, in the batch experimental mode, of the adsorption of lysozyme on the dye-affinity sorbents varied from 20 s for the non-porous glass beads with a size range of 20-40 microns to more than 60 min in the case of a porous sorbent with a particle diameter of 100-300 microns and 60 nm pore size. Furthermore, equilibration times, which represent the overall adsorption rates incorporating all the non-equilibrium effects, increased with all affinity systems when adsorption took place in the non-linear portion of the isotherm. The most dramatic increase was observed when sorbents with relatively high protein size to pore size ratios, lambda, were employed.
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
F B
FB
Anspach
A
A
Johnston
H J
HJ
Wirth
K K
KK
Unger
M T
MT
Hearn
D000426:Alcohol Dehydrogenase / Q000032:analysis; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity; D004396:Coloring Agents; D004058:Diffusion; D006801:Humans; D007700:Kinetics; D009113:Muramidase / Q000032:analysis; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D012709:Serum Albumin / Q000032:analysis; D013816:Thermodynamics; D014227:Triazines
D000596:Amino Acids; D004396:Coloring Agents; D010455:Peptides; D011506:Proteins; D012709:Serum Albumin; D014227:Triazines; C012842:Cibacron Blue F 3GA; D000426:Alcohol Dehydrogenase; D009113:Muramidase