Mechanochemistry offers a unique opportunity to modify and manipulate crystal forms, often providing new products as compared with conventional solution methods. While promising, there is little known about how to...
A novel stationary phase for affinity separations is presented. This material is based on sintered borosilicate glass readily available as semi-finished filter plates with defined porosity and surface area. The material shows fast binding kinetics and excellent long-term stability under real application conditions due to lacking macropores and high mechanical rigidity. The glass surface can be easily modified with standard organosilane chemistry to immobilize selective binders or other molecules used for biointeraction. In this paper, the manufacturing of the columns and their respective column holders by 3D printing is shown in detail. The model system protein A/IgG was chosen as an example to examine the properties of such monolithic columns under realistic application conditions. Several specifications, such as (dynamic) IgG capacity, pressure stability, long-term performance, productivity, non-specific binding, and peak shape, are presented. It could be shown that due to the very high separation speed, 250 mg antibody per hour and column can be collected, which surpasses the productivity of most standard columns of the same size. The total IgG capacity of the shown columns is around 4 mg (5.5 mg/mL), which is sufficient for most tasks in research laboratories. The cycle time of an IgG separation can be less than 1 min. Due to the glass material’s excellent pressure resistance, these columns are compatible with standard HPLC systems. This is usually not the case with standard affinity columns, limited to manual use or application in low-pressure systems. The use of a standard HPLC system also improves the ability for automation, which enables the purification of hundreds of cell supernatants in one day. The sharp peak shape of the elution leads to an enrichment effect, which might increase the concentration of IgG by a factor of 3. The final concentration of IgG can be around 7.5 mg/mL without the need for an additional nanofiltration step. The purity of the IgG was > 95% in one step and nearly 99% with a second polishing run.
One of the challenges of high-temperature polymer electrolyte
membrane
fuel cells is the poisoning of the Pt catalyst with H3PO4. H3PO4 is imbibed into the routinely
used polybenzimidazole-based membranes, which facilitate proton conductivity
in the temperature range of 120–200 °C. However, when
leached out of the membrane by water produced during operation, H3PO4 adsorbs on the Pt catalyst surface, blocking
the active sites and hindering the oxygen reduction reaction (ORR).
The reduction of H3PO4 to H3PO3, which occurs at the anode due to a combination of a low
potential and the presence of gaseous H2, has been investigated
as an additional important contributing factor to the observed poisoning
effect. H3PO3 has an affinity toward adsorption
on Pt surfaces even greater than that of H2PO4
–. In this work, we investigated the poisoning
effect of both H3PO3 and H3PO4 using a half-cell setup with a gas diffusion electrode under
ambient conditions. By means of in situ X-ray absorption spectroscopy,
it was possible to follow the signature of different species adsorbed
on the Pt nanoparticle catalyst (H, O, H2PO4
–, and H3PO3) at different
potentials under ORR conditions in various electrolytes (HClO4, H3PO4, and H3PO3). It was found that H3PO3 adsorbs in a pyramidal
configuration P(OH)3 through a Pt–P bond. The competition
between H3PO4 and H3PO3 adsorption was studied, which should allow for a better understanding
of the catalyst poisoning mechanism and thus assist in the development
of strategies to mitigate this phenomenon in the future by minimizing
H3PO3 generation by, for example, improved catalyst
design or adapted operation conditions or changes in the electrolyte
composition.
In this work (Part II) the application of new polystyrene based microcarriers in cell culture technology is demonstrated. Carriers with a variety of surface modifications were tested as a growth support for cell line BHK 21. The growth behavior of the cells and cell to surface attachment were compared to Cytodex 3 (Pharmacia), which was used as a reference carrier. To select carriers with growth supporting surfaces, broad screening in petri dish experiments was carried out. Candidates with the highest growth rates were investigated in spinner flask experiments in further detail. Polystyrene carrier with a surface modification like triethylamine, maltamine or N-methylglucosamine were able to support growth as good or better as the reference carrier Cytodex 3. Economies of ingredients and ease in laboratory handling could make amine-modified polystyrenes a competitive alternative to currently commercially available microcarrier types.
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