PMC

Schematic overview of the processes investigated in this study. The Reference process is shown at the top, consisting of centrifugation-based precipitation, wash, and re-dissolution. Process transfer to a cross-flow filtration (CFF) unit resulted in the Basic process. Transferred process steps are wash and re-dissolution, highlighted in blue and green, respectively. Wash and re-dissolution are multiple process steps consisting of repeated centrifugation (highlighted in yellow) in the Reference process. In the Basic process, these are reduced to two consecutive diafiltration (DF) steps by simply switching between diafiltration buffers (). Alternative CFF process variants, either Nuclease or mmSEC, are modifications from the Basic process. The Nuclease process adds a nucleic acid digestion and a 300 kDa wash step preceding precipitation and continues like the Basic process. The mmSEC process sequence is identical to the Basic process sequence but has a modified re-dissolution step (DF II) including a multimodal size-exclusion chromatography (mmSEC) column in the permeate line. (NH₄)₂SO₄, ammonium sulfate; HCP, host cell protein; UF/DF, ultrafiltration/diafiltration; VLP, virus-like particle.

Performance of P_GTH1 ‐D1240 in basic and optimized fed‐batch cultivations. A schematic overview of a basic constant glucose‐based process for P_GTH1 (a) and a typical three‐stage methanol‐driven process (b) for P_AOX1 is shown. Production kinetics (specific productivity q_P at different specific growth rates) from fed‐batch cultivations (c) led to the design of an optimized feeding strategy for P_GTH1 ‐D1240 (d). The optimized process resulted in increased final supernatant titers and volumetric productivity (Q_P) for P_GTH1 ‐D1240 which was even outperforming the P_AOX1‐driven process (Table 3 and e, f: the basic P_GTH1 ‐D1240 process (blue), the P_GTH1‐optimized process (green) and the standard P_AOX1 process (gray) [Color figure can be viewed at wileyonlinelibrary.com]

Basic process of T-lymphocyte activation by exposure to an APC and IFN-γ . A number of cytokines are involved in the activation process

The diagram of the basic locating point of the spinous process locating method. (A) The central vertical line of zygapophysial joint. (B) The horizontal line of superior margin root of spinous process. Point A = the basic locating point.