SUMMARY

Biomanufacturing for advanced therapies depends on the reliable and reproducible growth of cells. Cells continuously secrete biomolecules that serve as the critical quality attributes (CQAs) for manufacturing, but current sensor technologies are incapable of characterizing these biomarkers in real-time. The Dynamic Sampling Platform (DSP) is a technology for real-time analysis of the chemical content of bioreactors. It synergistically combines aseptic, spatially resolved, direct-from-reactor sampling with inline sample treatment and rapid electrospray ionization mass spectrometry (ESI-MS) sensing to fill the void of existing process analytical technologies for real-time CQA monitoring in biomanufacturing. DSP samples non-invasively from near where cells are growing to capture the rich microenvironment, with high relative concentrations of biomarkers, and subsequently treats these samples for real-time electrospray ionization mass spectrometry (ESI-MS) analysis. ESI-MS is a powerful analytical technology for the detection of biomolecules, but it is hindered by a need for extensive sample treatment which has made the technique largely an offline approach. The DSP utilizes a novel “active sample treatment” to prepare complex biochemical samples for ESI-MS by conditioning the sample via selective salt removal, MS enhancing chemical infusion, and biomolecule retention. The DSP is produced by employing advanced microfabrication techniques. The final device utilizes a micro-mass exchanger which replaces time consuming sample preparation steps with a ~1 second flow through reactor. The results of this dissertation demonstrate the DSP's utility to help unlock the potential of advanced therapeutic cell manufacturing by providing detailed biochemical information about the process for closed feedback control of bioreactors, allowing for nascent therapies to reach a broad patient population. https://bluejeans.com/318315274