Tangential flow filtration (TFF) is used in the biotechnology industry for concentrating the therapeutic proteins and exchanging it into the formulation buffer to produce bulk drug substance (BDS). The TFF unit operation is typically optimized based on a target processing time and protein concentration. The quality of the resulting protein is often a function of TFF process parameters, such as crossflow velocity and BDS concentration (short term stability), and formulation buffer conditions (long term stability). Protein stability and quality are often evaluated in the UF buffer (buffer from the previous step) and the DF buffer (formulation buffer). This work is focused on the impact of the buffer exchange process on the quality of the intermediate protein pool. To examine the effect of process buffers on the protein during buffer exchange, protein solubility as a function of protein concentration and buffer composition was evaluated by high-throughput membrane plates. The high throughput results were compared to in-process samples from operating TFF using cassettes. Furthermore, the cassettes with different screen configurations were evaluated for additional impact of shear stress on the protein quality. Experimental results were obtained using monoclonal antibodies and fusion proteins that demonstrate different levels of stability during normal TFF processing. Protein quality was evaluated using micro-flow imaging, dynamic light-scattering, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and size-exclusion chromatography (SEC) as a function of buffer composition and protein concentration. An optimal window of operation for the TFF process is identified for future evaluation.

Impact of buffer exchange and protein concentration on biologics quality during ultrafiltration and diafiltration (UF/DF)

251st National Meeting (2016)

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