Development of an Online Monitoring and Sampling Scheme for Recombinant Protein Production

University essay from Lunds universitet/Bioteknik (master)

Abstract: During the past few years the focus on bioprocess monitoring and control has increased. An area still under intensive development is the sampling methods for efficient and rapid analysis during fermentations. In this thesis, an online-sampling and monitoring scheme was developed using the versatile automated continuous flow system (VersAFlo) for recombinant protein production. From the implementation of the scheme with a fermentation, recovery of intra-cellular protein occurred within 19 minutes. The binding and washing buffer were determined to be efficient at 30 mM imidazole. A second studied aspect, cell-lysis using the VersAFlo, resulted in observed trends on microliter scale (1:1 and 1:2 cells to BugBuster Master Mix ® and time intervals 0, 30, 60 120, 240 seconds). Increasing the ratio and prolonging the reaction time resulted in greater degree of lysate whilst remaining on a scale consisting of microliters and seconds. Additionally, a dilution translation tool was created for optical density measurements at 620 nm using the system set-up. A strong correlation was observed between the translation tool and external online OD measurements from the fermentations. All studies build on the creation of running schemes which are broken down into step-by-step guidelines.

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