Background

HJB’s Highly Intensified Continuous Bioprocessing (HiCB) platform is ideally suited for producing a wide range of molecules, including monoclonal antibodies, multi-specific antibodies, and recombinant proteins, regardless of their stability.  

Results

Significant productivity increases with “Plug and Play” perfusion (same cell lines in FB vs. Perfusion)

By using the same cell lines and in-house developed chemically defined cell culture media, our “Plug and Play” continuous perfusion process has demonstrated significant improvements over the conventional fed-batch process, even without media or process optimization.  It achieves an average of over a 50% increase in cell-specific production rates and delivers at least an 8-folds increase in annual output. “Plug and Play” process is typically employed in first-in-human (FIH) manufacturing to ensure high quality production and speed to clinic.

Significant upward potential via “Push to High” strategy

To further enhance productivity in preparation for late-stage clinical development and commercial launch, the FIH “Plug and Play” perfusion process is improved through media and process optimization. This is followed by increases in perfusion rate (reactor volume per day or RV/day) to achieve higher cell densities and productivities, a strategy referred to as “Push to High”. An example of “Push to High” is shown below for Molecule E, an IgG 4, where step increases in perfusion rate from 1.5 RV/day to 2.25 RV/day, and subsequently to 2.5 RV/day, resulted in up to 100% increase in volumetric productivities. The average specific production rate during these changes was 52 pg/cell-day, which is double that of the fed-batch process. Notably, these improvements were achieved without media or process optimization; with such optimizations, the productivity gains would be even greater.

Significant potential of “Push to High” strategy for our continuous perfusion platform is further demonstrated for five molecules, including monoclonal and bi-specific antibodies, where volumetric productivities were increased up to 8 g/L-day, without media or process optimization.  At such high productivity, a single 500-liter single-use bioreactor can produce over 70 kg of drug substance per 30-day perfusion run, equating to an annual output of > 750 Kg.

Continuous perfusion enhances control of product quality

Another key advantage of continuous perfusion is its ability to control the extracellular environment, including parameters such as pH, temperature, DO, pCO2, nutrient levels, waste metabolites, and osmolality.  By maintaining these factors in a controlled state, continuous perfusion helps ensure a consistent metabolic state.  Combined with significant decreases in product residence time in the bioreactor and high culture viabilities, continuous perfusion further enhances control over product quality, ensuring consistent and high product quality throughout the production process.

•AstraZeneca (2020) Perfusion reduces bsAb aggregation via mitigating mitochondrial dysfunction‑induced GSH oxidation and ER stress in CHO cells.  Sci. Reports. 6;10(1):16620 

•Amgen 2020 Improving product quality and productivity of bispecific molecules through the application of continuous perfusion principles, Biotech Progress. 36(4):e2973 

•Sanofi (2019) Perfusion Cell Culture Decreases Process and Product Heterogeneity in a Head-to-Head Comparison With Fed-Batch.  Biotechnology J. Feb;14(2):e1700733 

•Amgen (2019) Perfusion CHO cell culture applied to lower aggregation and increase volumetric productivity for a bispecific recombinant protein.  J. Biotech.  304:70-77 

•Merck (2019) Perfusion cell culture for the prod’n of conjugated recomb. fusion proteins reduces clipping and quality heterogeneity compared to batch mode processes.  J. Biotech. 302:26-31