Effect of controlled redox potential and dissolved oxygen on the in vitro refolding of an E. coli alkaline phosphatase and chicken lysozyme

Meneses‐Acosta, Angélica; Vizcaíno-Meza, Luis Rodolfo; Ayala-Castro, Hector G.; Contreras, Martha A.; Ortega‐López, Jaime; Ramı́rez, Octavio T.

doi:10.1016/j.enzmictec.2013.03.008

Cited by 3 publications

(1 citation statement)

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“…The progress of the refolding operation can be captured by measuring changes in physical variables such as ORP, pH, and temperature. ORP is an indicator of refolding as redox conditions vary widely during solubilisation and refolding (Meneses‐Acosta et al, ; Shultz, Hart, & Keener, ). Solubilisation requires reducing conditions to unfold nonnative intramolecular and intermolecular disulphide bonds while keeping cysteines in their reduced state.…”

Section: Resultsmentioning

confidence: 99%

Process analytical technology implementation for protein refolding: GCSF as a case study

Hebbi

Thakur

Rathore

2019

Biotech & Bioengineering

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Process analytical technology is gaining interest in the biopharmaceutical industry as a means to enable consistency in processing and thereby in product quality via process control. Protein refolding is known to be significantly impacted by critical process parameters and feed material attributes including composition and pH of the solubilisation and refolding buffers. Hence, to achieve robust process control and product quality, these attributes and parameters need to be monitored. This paper presents an approach towards statistical process control and monitoring of protein refolding, from buffer preparation to refold quenching, during manufacturing of therapeutic proteins from Escherichia coli based systems. The proposed approach utilises measurements of online redox potential, temperature, and pH for development of a statistical model. The model has then been integrated with LabView to permit real‐time monitoring of the refolding process. The proposed system has been demonstrated to successfully identify process deviations and thereby enable process control for manufacturing product of consistent quality.

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Section: Resultsmentioning

confidence: 99%