Enabling Bioprocessing 4.0 with Culture Biosciences and 908 Devices

Bioprocessing 4.0 aims to leverage advanced digital technologies, automation, and data-driven approaches to transform the way biopharmaceuticals are produced. Culture Biosciences is at the leading edge of this revolution by offering clients access to cloud-connected bioreactors and data science capabilities that enable the digital transformation of biomanufacturing processes for Process Development (PD). Culture Biosciences’ bioreactors autonomously capture and integrate real-time data, via cloud-enabled controllers which can be monitored live or analyzed using Culture’s Console™. Culture’s real-time data analysis and data driven modeling capabilities, working in conjunction with novel Process Analytical Technologies (PAT), enable clients to accelerate their biomanufacturing PD work. 

In an effort to enable innovative Bioprocessing 4.0 PD workflows using PAT methods, Culture Biosciences and 908 Devices formed a collaboration to test MAVERICK™ – a novel Raman spectroscopy powered in-line PAT – in Culture Biosciences’ cloud-connected 250mL bioreactors. MAVERICK provides real-time, in-line bioprocess analysis and control, right out of the box without the costs, complexities, and risks of conventional Raman approaches. The vertically integrated hardware stack at Culture Biosciences enables rapid prototyping and modification of our proprietary bioreactors to support a client’s process-specific PAT needs. Culture Biosciences’ R&D - Hardware Team rapidly fitted a patch dissolved oxygen (DO) probe and configured a PAT port for the MAVERICK in-line immersion probe, unlocking a new capability for our clients interested in using real-time in-line Raman PAT to monitor and control their PD bioprocesses.

(left) View of Culture Biosciences Cloud Bioreactor Lab with two bioreactors (288 and 289) setup with 908 Devices MAVERICK™ PAT. (right) zoomed in view of the in-line probe inserted in a 250 mL vessel.

To test the cloud-connected biomanufacturing process with PAT capabilities, Chinese Hamster Ovary (CHO) mammalian cells were cultured in four 250mL bioreactors using a standard fed-batch cell culture protocol. Two bioreactors were fitted with MAVERICK™ in-line immersion PAT probes supplied by 908 Devices and two bioreactors were used as baseline experimental controls. MAVERICK measured glucose, lactate, and biomass in real-time continuously throughout the culture duration. Feed media, glucose and other nutrients were supplemented to the bioreactors following a fully-automated protocol, controlled by Culture Biosciences’ cloud-connected  proprietary controller and monitoring technology. Nutrient feeding steps can be activated manually or automatically, depending on the application and client process need. In the protocol tested, supplemental feeding was activated using feedback from daily off-line measurements to adjust glucose levels in the bioreactor. Bioreactors were aseptically sampled at 24h intervals and analyzed for glucose, lactate, and other metabolites (Flex2) and cell density (Vi-CELL BLU). Following experiment completion (14 days), the data from the off-line and in-line devices was analyzed using Culture’s Console™ software and Application Programming Interface (API) to assess probe performance within Culture’s 250mL bioreactor systems.

Glucose measurements show alignment between off-line data from Flex2 and in-line PAT values from the MAVERICK™ system (RMSE = 0.32 g/L). The figure below suggests that the MAVERICK PAT measurements were accurately reflecting the live culture conditions within the bioreactors. In-line PAT measurements can track the effect of feed and media addition events (blue line) in real-time within the bioreactor environment. These dynamic cell culture condition changes are not captured in the off-line daily sample measurements (orange line).

Glucose measurements in a Culture Biosciences bioreactor, in-line by MAVERICK™ PAT (blue) and daily off-line measurements by Flex2 (orange).

Culture Biosciences’ bioreactors enable accurate volume monitoring through cloud-connected sensors and controllers in conjunction with a data aggregation backend. The real-time monitoring of this data enables accurate tracking of all volume changing events in the bioreactor throughout  the time-course experiment. From the generated feed volume data set, feed additions explain the increase in glucose concentration seen with in-line MAVERICK PAT measurements.  

Glucose spikes measured by MAVERICK coincide with feed pump data captured and processed by Culture Biosciences’ cloud bioreactors.

Similar to the observed glucose measurements, lactate levels are accurately measured using the MAVERICK™ probe when compared to off-line analytics  (RMSE = 0.29 g/L).  

Lactate measured in-line by PAT tool (blue) and through daily off-line samples (orange). 

The MAVERICK™ PAT also provides biomass measurements; these values were compared to the off-line Vi-CELL BLU total cell density. Raman based technologies typically require a calibration curve for a conversion into physical units. The figure below shows the raw signal measurements from the MAVERICK (PAT) (in biomass units, BMU) on the left side y-axis and Vi-CELL BLU measured total cell density (in million cells/mL) on the right side y-axis. The figure shows that the two curves are in agreement, indicating that MAVERICK is capable of effectively capturing the underlying physical signal. A divergence in the two measurements occurs beyond 250 hours, which can be attributed to the reduction of viable cells that occurs at the end of a typical mammalian cell culture experiment. The reduction in cell viability results in increased cell debris which may be a confounding variable for  in-line PAT measurements, while the Vi-CELL BLU’s visual measurements can be tuned to exclude measuring lysed cells. 

Biomass (arbitrary units)  measured in-line by MAVERICK™ PAT (blue, left y-axis) compared against total cell density (million cells/mL) available from daily off-line measurements (orange, right y-axis). 

In summary, these experiments  show how the flexibility and vertical integration of Culture’s bioreactor platform allows for the rapid integration of PAT probes into 250mL bioreactors. The data generated utilizing Culture Bioscience’s bioreactors, cloud-based controllers and software Console™ together with MAVERICK™, a novel Raman spectroscopy powered in-line PAT, was found to track closely with off-line measurements for multiple critical analytes. 

PAT in bioreactors can accelerate your understanding by providing in-line, continuous data about cell culture conditions. Continuous data streams are useful in precise monitoring and control of your bioprocess. Culture Biosciences is at the leading edge of Bioprocessing 4.0 by offering clients access to cloud-connected bioreactors and data science capabilities that enable the digital transformation of biomanufacturing processes.

If you are interested in accelerating bioprocess development towards Bioprocessing 4.0 with advanced digital technologies, automation and data-driven approaches to transform the way biopharmaceuticals are produced, please reach out: inquiries@culturebiosciences.com