FIGURES
Figure 1 : The µSI process analyzer system for MAM sample
preparation. (a) The fluidic diagram, (b) Left : The µSI process
analyzer system connected to a LC-MS system on the manufacturing floor.Right : Close up view of the upper compartment inside the
functionalized µSI process analyzer system.
Figure 2 : The µSI process analyzer system for glycosylation
profile assay sample preparation. (a) The fluidic diagram, (b)Left : The µSI process analyzer system connected to a Waters
Acquity UPLC system in development lab. Right : Close up to the
upper compartment inside the functionalized µSI process analyzer system.
Figure 3 : The A2P2 system. (a) The fluidic diagram, (b) A
configured A2P2 system utilizes a commercially available LC system. All
components are integrated vertically for efficient use of laboratory
footage. (c) The sample preparation unit operation beneath the
autosampler in the A2P2 system. (RxN Coil: reaction coil)
Figure 4 : Engineering design of the FTP and its holder. (a)Left : FTV isometric view, cross-section of vial with arrows
indicating liquid flow. Middle : FTV holder. Right : Full
assembly of vial holder and FTV. (b) The FTV is a key part for the
breadth of functionalities in the A2P2 system. The extended gutter and
added lip to help prevent surface tension. Extrusion at the end of the
gutter is conveniently placed directly over the waste port.
Figure 5 : The logical process developed by utilizing the LC
chromatography data system software to enable autonomous operation in
the A2P2 system. The uses of SST and IRC workflows enable autonomous and
self-directed sample preparation and/or assay execution. (LLOQ: lower
limit of quantitation, ULOQ: upper limit of quantitation, SEC: size
exclusion, SST: system suitability testing, IRC: intelligent run
control)
Figure 6 : (a) The TIC readout from the LC-MS connected µSI
process analyzer system demonstrated good robustness over the course of
6 days continuous reference standard processing. (b) The real-time
product quality output from a production bioreactor. The trending curves
for deamidation and clips generated from the real-time MAM results can
be used for process decisions, in-process control, and product release.
Figure 7 : (a) Glycan chromatogram generated from a reference
standard processed by the integrated HILIC UPLC µSI process analyzer,
full and zoomed view. (b) The performance of the system was verified by
daily processing of the reference standard for 32 days. (c) Continuous
glycan monitoring of a production bioreactor by the system over the
duration of entire upstream bioproduction process.
Figure 8 : Capabilities of the A2P2 system. (a) Aggregation
assessment for biological products, (b) Charge variant profile for
biological products, (c) Glycosylation profile for biological products,
(d) Multi-attribute method for biological products. Out-of-scope flow
paths were masked for simplicity.