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Based on the folding kinetic the dynamic behavior of such a continuous refolding reactor was simulated under different operating conditions.
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A novel coiled flow inverter (CFI) based plug flow reactor has been developed for continuous refolding of granulocyte colony stimulating factor (GCSF), a biotech therapeutic product.
To overcome the shortcomings of batch chromatography, the size-exclusion refolding reactor may be operated in a continuous mode, with the aid of Simulated Moving Bed (SMB) technology.
The performance of the continuous refolding protocol has been compared with an optimized batch refolding protocol.
With this continuous refolding process, we increased the refolding and cleavage yield of both model proteins by 10% compared to batch dilution refolding.
For experimental verification of the model a continuous refolding of a model protein (α-lactalbumin) was performed in a CSTR.
Based on the actual refolding data, we compared throughput, productivity, and buffer consumption between two batch dilution refolding processes – one using urea for IB dissolution, the other one using NaOH for IB dissolution – and our continuous refolding process.
A new continuous refolding method using a four-zone simulated moving bed process based on the size exclusion mechanism was developed in order to overcome the disadvantages associated with inclusion body refolding in a batch dilution and chromatography.
The higher complexity of the continuous refolding process was rewarded with higher throughput and productivity as well as significantly lower buffer consumption compared to the batch dilution refolding processes.
Therefore, in this work we present a methodology using a SEC refolding reactor model, supported by a library of reaction mechanisms, to estimate a suitable reaction scheme using experimental SEC refolding data.
Accordingly, the comparison revealed the significant advantages that SMB technology has to offer to the design of chromatographic refolding reactors.
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