Abstract
Soluble expression of recombinant proteins in E. coli is often done by translocation of the product across the inner membrane (IM) into the periplasm, where it is retained by the outer membrane (OM). While the integrity of the IM is strongly coupled to viability and impurity release, a decrease in OM integrity (corresponding to increased “leakiness”) leads to accumulation of product in the extracellular space, strongly impacting the downstream process. Whether leakiness is desired or not, differential monitoring and control of IM and OM integrity are necessary for an efficient E. coli bioprocess in compliance with the guidelines of Quality by Design and Process Analytical Technology. In this review, we give an overview of relevant monitoring tools, summarize the research on factors affecting E. coli membrane integrity and provide a brief discussion on how the available monitoring technology can be implemented in real-time control of E. coli cultivations.
Highlights
Despite lockdowns in China, SARS-CoV-2 spread rapidly throughout the world by predicted routes of transportation (Bogoch et al, 2020) and spread faster than any other emerging infectious disease in recent decades (Angelo et al, 2019; Halstead & Wilder-Smith, 2019; Tuite et al, 2019; Wilder-Smith, Chang, & Leong, 2018)
The testing strategy efficacies as a function of the total number of tests and corresponding costs in US dollars are shown in Fig. 3(a)e(b), respectively, for different times ti, the strategy started to be applied
The testing strategy efficacies as a function of the total number of tests and corresponding costs in US dollars are shown in Fig. 4(a) and (b), respectively, for different dates of start
Summary
Despite lockdowns in China, SARS-CoV-2 spread rapidly throughout the world by predicted routes of transportation (Bogoch et al, 2020) and spread faster than any other emerging infectious disease in recent decades (Angelo et al, 2019; Halstead & Wilder-Smith, 2019; Tuite et al, 2019; Wilder-Smith, Chang, & Leong, 2018). The first case of COVID-19 in Latin America was confirmed on February 26, 2020, in the Sa~o Paulo metropolis, the most populous city in the Southern hemisphere (~12 million people) (Instituto Brasileiro de Geografia e Estatística, 2020). Travel reports and subsequent genetic analyses confirmed that the first detected infection was acquired via importation of the virus from Northern Italy (Candido et al, 2020; Jesus et al, 2020). Brazil has reported the largest number of cases in Latin America (4,238,446 reported cases and 129,522 deaths as of September 11, 2020) (Ministerio da Saúde do Brasil, 2020). SARS-CoV-2 spread rapidly within Brazil (Carmo et al, 2020), often associated with urban centers with social deprivation (Souza et al, 2020) and has been detected in the majority of the 27 federal states of Brazil. Using transmission pairs of SARS-CoV-2 reported to the Brazilian Ministry of Health, the mean and standard deviation for the serial interval was estimated to be 2.97 and 3.29 days respectively (Prete et al, 2020)
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