Recovery of antibodies with Protein A affinity chromatography columns has become the standard for the biotechnology industry. Membrane affinity chromatography has not yet experienced extensive application due to the lower capacity of membrane supports compared to chromatographic beads. In this work, new affinity membranes endowed with an interesting binding capacity for human IgG are studied in view of their application in the capturing step of a monoclonal antibody production process. The membranes have been extensively tested with pure IgG solutions and with a cell culture supernatant containing IgG 1 . The effects of feed flow rate and IgG concentration on the separation performances have been studied in detail, considering in particular binding capacity, selectivity and recovery. These new high capacity affinity membranes appear good candidates to avoid the throughput limitations and other well-known drawbacks of traditional bead-based chromatographic columns.
Highly porous particles with internal triply periodic minimal surfaces were investigated for sorption of proteins. The visualization of the complex ordered morphology requires complementary advanced methods of electron microscopy for 3D imaging, instead of a simple 2D projection: transmission electron microscopy (TEM) tomography, slice-and-view focused ion beam (FIB) and serial block face (SBF) scanning electron microscopy (SEM). The capability of each method of 3D image reconstruction was demonstrated and their potential of application to other synthetic polymeric systems was discussed. TEM has high resolution for details even smaller than 1 nm, but the imaged volume is relatively restricted (2.5 μm)3. The samples are pre-sliced in an ultramicrotome. FIB and SBF are coupled to a SEM. The sample sectioning is done in situ, respectively by an ion beam or an ultramicrotome, SBF, a method so far mostly applied only to biological systems, was particularly highly informative to reproduce the ordered morphology of block copolymer particles with 32–54 nm nanopores and sampling volume (20 μm)3.
We report the preparation of crosslinked membranes for organophilic filtration, by reacting a new polytriazole with free OH groups, using non-toxic poly (ethylene glycol) diglycidyl ether (PEGDE). The OH-functionalized polymer was obtained by converting the oxadiazole to triazole rings with high yield (98%). The maximum degree of crosslinking is achieved after 6 h of reaction. The crosslinked polytriazole membranes are stable in a wide range of organic solvents and show high creep recovery, indicating the robustness of crosslinked membranes. The influence of different casting solutions and different crosslinking time on the membrane morphology and membrane performance was investigated. The membranes performance was studied in dimethylformamide (DMF) and (tetrahydrofuran) THF. We achieved a permeance for THF of 49 L m-2 h-1 bar-1 for membranes with molecular weight cut off (MWCO) of 7 kg mol-1 and a permeance for THF of 17.5 L m-2 h-1 bar-1 for membranes with the pore dimensions of the membranes to have a MWCO, which covers ultra-and nanofiltration range.
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