Generation of equally sized particle plaques using solid-liquid suspensions.
A device is presented for the generation of equally sized plaques of sensitive particles in a 96-well format. The resulting particle plaques can be used for the measurement of adsorption isotherms and uptake kinetics in protein chromatography or for immobilization reactions. The particle plaques are formed from suspensions with a vacuum device that is designed as a reusable sandwich module. The particles are retained by a mesh while the solvent is removed by the vacuum. As most particles used for protein chromatography are sensitive to mechanical stress and dehydration, the vacuum device is gentle enough to allow the use of these particles, thus eliminating the uncertainty of slurry preparation and pipetting. Apparatus characteristics and preparation procedures are described precisely. The physical intactness of the particles after the preparation procedure is proved by microscopic analysis. Data on the uniformity of the obtained resin plaques with respect to the reproducibility of their adsorption performance is given. Finally, adsorption isothermal and kinetic data of BSA on an ordinary HIC system obtained by high-throughput measurements are shown as an application example.
Herrmann
Tim
T
Institute of Biotechnology, Research Center Juelich, 52425 Juelich, Germany.
Schröder
Magnus
M
Hubbuch
Jürgen
J
D000327:Adsorption; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D007103:Immobilization; D007700:Kinetics; D010316:Particle Size; D011506:Proteins / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013816:Thermodynamics
D011506:Proteins; D012997:Solvents