Theoretical investigation of the optimum particle size for the resolution of proteins by size-exclusion chromatography

摘要: Abstract A theoretical discussion is presented of the possibilities dramatic acceleration size-exclusion chromatographic separation proteinsas a result marked decrease in diameter gel particles used to pack column. The use very small particles, range 1–2 μm, made compatible with moderate inlet pressures by requirement low mobile phase velocity, due diffusion coefficient high-molecular-weight compounds. Thus, conditions regarding (i) suppression eluent flow intra-particle pores, (ii) avoidance shear degradation proteins because excessive viscous stress and (iii) size exclusion protein molecules from some interstitial channels can be met for molecular weights up several hundred thousand. optimum particle discussed, it shown that efficiencies few tens thousands plates, permitting peak capacities 30–40, could generated using 20–30 cm long columns packed 1- or 2-μm particles. analysis time depends on weight but, these columns, would order 1 h. results this are supported experimental data, illustrating practical approach.