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  • Performance Comparison of a Laterally-Fed Membrane Chromatography (LFMC) Device with a Commercial Resin Packed Column.

Performance Comparison of a Laterally-Fed Membrane Chromatography (LFMC) Device with a Commercial Resin Packed Column.

Membranes (2019-11-02)
Pedram Madadkar, Rahul Sadavarte, Raja Ghosh
ABSTRACT

The use of conventional membrane adsorbers such as radial flow devices is largely restricted to flow-through applications, such as virus and endotoxin removal, as they fail to give acceptable resolution in bind-and-elute separations. Laterally-fed membrane chromatography or LFMC devices have been specifically developed to combine high-speed with high-resolution. In this study, an LFMC device containing a stack of strong cation exchange membranes was compared with an equivalent resin packed column. Preliminary characterization experiments showed that the LFMC device had a significantly greater number of theoretical plates per metre than the column. These devices were used to separate a ternary model protein mixture consisting of ovalbumin, conalbumin and lysozyme. The resolution obtained with the LFMC device was better than that obtained with the column. For instance, the LFMC device could resolve lysozyme dimer from lysozyme monomer, which was not possible using the column. In addition, the LFMC device could be operated at lower pressure and at significantly higher flow rates. The devices were then compared based on an application case study, i.e., preparative separation of monoclonal antibody charge variants. The LFMC device gave significantly better separation of these variants than the column.

MATERIALI
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Sigma-Aldrich
Lisozima, lyophilized powder, protein ≥90 %, ≥40,000 units/mg protein
Sigma-Aldrich
Albumina, lyophilized powder, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Sodio citrato tribasico, ACS reagent, ≥99.0%
Sigma-Aldrich
Sodio cloruro, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Acido citrico, 99%
Sigma-Aldrich
Sodium phosphate monobasic, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Conalbumin from chicken egg white, BioReagent, suitable for cell culture