Specific and Nonspecific Adsorption in Affinity Chromatography. Part II. Kinetic and Mass Transfer Studies

• Published in: Industrial & engineering chemistry research 2001-01, Vol.40 (1), p.377-383
• Main Authors: MARTIN DEL VALLE, E. M, GALAN, M. A
• Format: Article
• Language: English
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Fundamental and applied biological sciences. Psychology
• ISSN: 0888-5885; 1520-5045
• DOI: 10.1021/ie000402f

The partition coefficient, adsorption equilibrium inside particles, adsorption kinetics, and mass transfer effects have been studied for asparaginase on Sepharose 4B activated with cyanogen bromide and with hexamethylenediamine and l-(+)-chlorosuccinamic acid as the spacer arm and ligand, respectively, for temperature and pH ranges of 298−302 K and 7.5−8.6, respectively, and for an ionic strength value of 0.05 M NaCl. A dynamic model has been developed to describe the adsorption. This model allowed us to obtain values of the diffusion coefficients and the forward surface interaction rate constants (D e ≈ 10-8 cm2/s, k 1 ≈ 10-4 mL mg-1 s-1). The forward surface interaction rate constant does not vary with the pH; however, an Arrhenius-type variation was observed with temperature. The values obtained for these constants range between 3.78 × 10-4 and 6.80 × 10-4 mL mg-1 s-1. The diffusion coefficient increases with pH (7.5−8.6). Additionally, within the temperature range studied, this coefficient was seen to follow Arrhenius-type behavior, allowing for a determination of the activation energies of the diffusion process for the different pH values (7.95−16.03 kcal/mol).