PROJECT 2: Dynamic Sampling Platform (DSP) for Cell State Analysis and Dynamic Bioreactor Monitoring via in situ nanoESI-MS and FTIR/ATR-SEIRAS
Test-Beds Involved: All
Our focus is on development of the Dynamic Sampling Platform (DSP), coupled to multi-mode analytical sensors such as nanoelectrospray ionization mass spectrometry (nano-ESI MS) and Fourier Transform Infrared Spectroscopy (FTIR), to create a multi-functional research instrument for cell reactor characterization and cell health monitoring that can also be integrated directly into therapeutic cell manufacturing quality control approaches.
Aim 1: DSP design capable of capturing changes in cell behavior in domains smaller than a single cell and at a time scale of seconds through rapid online nano-ESI MS analysis made possible with a microfabricated mass exchanger, while lower abundance chemicals, or chemicals undergoing slower processes, are observed using parallel on-line high performance liquid chromatographic separation
Aim 2: Development and demonstration of the DSP microfabrication process: batch microfabrication enables reproducible production of DSPs with excellent control of critical feature dimension
Aim 3: Study the detection of IDO and its presence in MSC cell culture using the FTIR technique ATR-SEIRAS with (a) IDO at different concentrations in buffer, (b) IDO with concentrations of other small molecules found in cell culture such as L-Tryp, and (c) using cell supernatant from Test-Bed 1;
Aim 4: Development of a prototype package (microfabricated device housing with fluidic connections) and DSP bio-reactor sampling interface;
Aim 5: Characterization of DSP performance, in combination with label free nano-ESI MS and ATR-SEIRAS detection, for monitoring human mesenchymal stem cells (hMSCs) and MSC culture supernatant grown under different bioreactor conditions.