Characteristics of the adsorption of immunoglobulin M onto Q Sepharose Fast Flow ion-exchangers.
Measurement of adsorption breakthrough curves in packed beds has shown that the amounts and rates of uptake of immunoglobulin M (IgM) onto the commonly used anionic ion-exchanger Q Sepharose Fast Flow (based on 6% agarose) are severely limited as a result of the large molecular size of this adsorbate (RMM 950,000). A similar ion-exchanger based on a more porous 4% agarose, Q Sepharose 4 Fast Flow was evaluated as an alternative adsorbent for the purification of IgM. Equilibrium adsorption isotherms and the effective diffusivities of IgM within these two adsorbents were measured. Q-Sepharose 4 Fast Flow was found to have a maximum capacity for IgM 2.5 times greater than that of Q Sepharose 6 Fast Flow and the effective diffusivity of IgM was found to be between 6 and 7 times greater than with the latter material. Comparison of the breakthrough curves obtained for these adsorbents at a variety of flow velocities confirm that Q Sepharose 4 Fast Flow is a superior adsorbent for the capture and purification of large proteins.
Chase
H A
HA
Department of Chemical Engineering, University of Cambridge, UK.
Machielse
B
B
Naveh
D
D
D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001709:Biotechnology; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004058:Diffusion; D005069:Evaluation Studies as Topic; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification*; D012685:Sepharose; D012709:Serum Albumin / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D012709:Serum Albumin; D012685:Sepharose