CHU Wenning, LIN Dongqiang, YAO Shanjing. High-throughput screening methods for protein adsorption evaluation with microtiter filter plate[J]. CIESC Journal, 2017, (6): 2399-2406.
DOI:
CHU Wenning, LIN Dongqiang, YAO Shanjing. High-throughput screening methods for protein adsorption evaluation with microtiter filter plate[J]. CIESC Journal, 2017, (6): 2399-2406.DOI: 10.11949/j.issn.0438-1157.20161789.
High-throughput screening methods for protein adsorption evaluation with microtiter filter plate
Aiming at the process optimization of chromatographic separation
the high-throughput screening methods of protein adsorption with microtiter filter plate were established for initial resin screening
adsorption properties evaluation
adsorption isotherms and adsorption kinetics measurement
and adsorption and elution conditions optimization. Firstly
the operation parameters of 96-well filter plate were optimized
and with two ion exchangers and two mixed-mode resins as the model
the bovine serum albumin adsorption was investigated with different resins and liquid conditions by means of microtiter filer plate. The binding capacity maps were obtained and the appropriate adsorption-desorption conditions were determined. Further
the microtiter filter plate was applied to measure the adsorption isotherms and adsorption kinetics with four resins at specific adsorption conditions
and the adsorption parameters were obtained. Finally
the elution conditions were optimized using microtiter filter plate. The microscale results were comparable to that with packed-bed separation
indicating that the methods developed in the present work were practicable. The results demonstrated that the high-throughput process development of protein adsorption with microtiter filer plate was feasible and can be used to fast screen the best reins and liquid conditions. New methods showed the advantages of low resource consumption
large experimental fluxes
short developing period
wide application and high stability
which would be one new method for development of the protein separation process.
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