A 50 mg/mL aqueous solution of bovine hemoglobin is to be purified by size-exclusion chromatography using a 75 x 2.5 cm i.d. glass column packed with 75 μm (diameter) Toyopearl HW65C. Intraparticle diffusivity of hemoglobin is 3.0 x 10^-7 cm²/s.

Estimate the resolving power per unit thermodynamic driving force for this protein in this column at 0.4, 0.8, and 1.2 cm/minute. What resolution is expected from a protein of comparable diffusivity whose inclusion porosity is 118% of hemoglobin? What size column would be required to obtain R = 1, as in Figure 15.21?

Transcribed Image Text:

Normalized Concentration 30 20 10 FTTT 0.0 0.2 0.4 0.6 Normalized Bed Length (1/L) 0.8 N 1.0 Figure 15.21 Fluid concentrations c₁ and ₂ plotted for chromatographic separation of 1 and 2 where t = 0.9511, 812 = 0.05, and R₁2 = 1 at z/L = 1.