Abstract
Background: Heterogeneity in recombinant protein quality (e.g. glycosylation) can result from a) changes in cultivation conditions during a fermentation process, b) product degradation due to differing residence times of product molecules in the supernatant, c) heterogeneities in intracellular PTM (post transcriptional modification) machinery, or from a combination thereof. To minimize heterogeneity in product quality it is of importance to evaluate the impact of these different mechanisms. Therefore we developed and applied a protocol, which can be used for analysis of a spectrum of different recombinant proteins and production cell lines aiming to provide a tool that enables growth phase related product analytics, for monitoring variations of the product during different phases of a cultivation process. Important for growth phase specific product analysis is the possibility to separate recombinant proteins produced during different growth phases. Exchanging media without changing its composition regarding nutrient and waste products should result in growth characteristics comparable to a cultivation without media exchange.
Highlights
Heterogeneity in recombinant protein quality can result from a) changes in cultivation conditions during a fermentation process, b) product degradation due to differing residence times of product molecules in the supernatant, c) heterogeneities in intracellular PTM machinery, or from a combination thereof
We developed and applied a protocol, which can be used for analysis of a spectrum of different recombinant proteins and production cell lines aiming to provide a tool that enables growth phase related product analytics, for monitoring variations of the product during different phases of a cultivation process
For a production cell line which growth behavior differs highly from the host cell line the cultivation needs to be adapted, or supernatants from different fermentation phases have to be prepared in advance for proper utilization at time of medium exchange
Summary
Heterogeneity in recombinant protein quality (e.g. glycosylation) can result from a) changes in cultivation conditions during a fermentation process, b) product degradation due to differing residence times of product molecules in the supernatant, c) heterogeneities in intracellular PTM (post transcriptional modification) machinery, or from a combination thereof. We developed and applied a protocol, which can be used for analysis of a spectrum of different recombinant proteins and production cell lines aiming to provide a tool that enables growth phase related product analytics, for monitoring variations of the product during different phases of a cultivation process. Important for growth phase specific product analysis is the possibility to separate recombinant proteins produced during different growth phases. Exchanging media without changing its composition regarding nutrient and waste products should result in growth characteristics comparable to a cultivation without media exchange
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
