Structure and protein adsorption mechanisms of clean and fouled tentacle-type anion exchangers used in a monoclonal antibody polishing step.

Autor: Corbett R; Department of Chemical Engineering, University of Virginia, 102 Engineers' Way, Charlottesville, VA 22904, USA., Carta G, Iskra T, Gallo C, Godavarti R, Salm JR
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2013 Feb 22; Vol. 1278, pp. 116-25. Date of Electronic Publication: 2013 Jan 09.
DOI: 10.1016/j.chroma.2013.01.006
Abstrakt: The properties of Fractogel(®) EMD TMAE HiCap (M), a tentacle-type anion exchange resin used for a polishing step in a monoclonal antibody (mAb) purification process, were investigated for both virgin and used samples to determine the influence of process related fouling. Inverse size exclusion chromatography indicated a bimodal distribution of pore sizes consisting mostly of small pores, 4-5 nm in radius and likely associated with the grafted tentacles. Similar results were obtained for resin samples fouled by process use, indicating that the core structure of these particles is unchanged. Transmission electron micrographs showed that the resin backbone matrix has a microgranular structure. However, a dense skin layer, 0.2-0.5 μm thick, was also seen at the exterior surface of the fouled particles. The binding capacity attained for BSA after 90 min of contact was 165 ± 4 mg/mL for both virgin and fouled samples, close to the equilibrium capacity of 178 ± 2 mg/mL attained after 24h. On the other hand, the capacities attained at 90 min for the much larger thyroglobulin were only 90 ± 4 and 25 ± 2 mg/mL, respectively, for virgin and fouled samples. The BSA adsorption kinetics was also slower for the fouled resin, but much larger kinetic differences between virgin and fouled resin were seen for thyroglobulin. Based on the shape of intraparticle protein concentration profiles determined by confocal laser scanning microscopy (CLSM), the protein transport mechanism is consistent with solid diffusion for both virgin and fouled resin samples and proteins. However, transport is hindered by the foulant layer to a much greater extent for thyroglobulin as a result of its larger size. Additional measurements indicated that the foulant layer is consistent with mAb aggregates irreversibly bound at the particle exterior surface.
(Copyright © 2013 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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