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N-1 Perfusion Platform Development Using a Capacitance Probe for Biomanufacturing.

Emily S C Rittershaus1, Matthew S Rehmann1, Jianlin Xu1

  • 1Biologics Development, Global Product Development and Supply, Bristol Myers Squibb Company, Devens, MA 01434, USA.

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|April 21, 2022
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Summary

Implementing a process analytical technology (PAT) tool with capacitance probes for N-1 perfusion cultures significantly enhances monoclonal antibody (mAb) production by Chinese hamster ovary (CHO) cells. This method optimizes perfusion rates, reduces media use, and supports high-density cell cultures for intensified biomanufacturing.

Keywords:
Chinese hamster ovary (CHO)capacitancecell-specific perfusion rate (CSPR)in-line probemonoclonal antibody (mAb)perfusion N-1platformprocess analytical technologies (PAT)process intensificationscale-up

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Area of Science:

  • Biotechnology
  • Bioprocess Engineering
  • Cell Culture Technology

Background:

  • Fed-batch process intensification for monoclonal antibody (mAb) production using Chinese hamster ovary (CHO) cells can be improved by N-1 perfusion seed trains.
  • Higher viable cell density (VCD) in N-1 perfusion seeds leads to increased inoculation VCD in the subsequent N-stage fed-batch, resulting in shorter culture duration, higher peak VCD, or improved titers.

Purpose of the Study:

  • To integrate a process analytical technology (PAT) tool, specifically an in-line capacitance probe, into an N-1 perfusion platform for automated perfusion rate control.
  • To evaluate the correlation between capacitance measurements and offline viable cell density (VCD) and assess the impact of real-time VCD-based perfusion control on media consumption and cell growth.

Main Methods:

  • An in-line capacitance probe was used to monitor viable cell density (VCD) in real-time.
  • The perfusion rate was automatically adjusted based on capacitance measurements to maintain a target cell-specific perfusion rate (CSPR).
  • Capacitance data was validated against offline VCD measurements and used for scaling up N-1 perfusion processes.

Main Results:

  • Capacitance measurements showed a linear correlation with offline VCD up to 130 × 10^6 cells/mL.
  • Online control of perfusion rate using CSPR reduced media usage by approximately 25% compared to volume-specific perfusion rates without negatively impacting cell growth.
  • A platform CSPR of 0.04 nL/cell/day was established and successfully applied to six mAb-producing cell lines, enabling rapid growth and high viability in four of them.

Conclusions:

  • The integration of a capacitance-based PAT tool enables efficient and automated control of N-1 perfusion processes for high-density CHO cell cultures.
  • This approach facilitates process development and intensification for fed-batch mAb production, leading to reduced resource consumption and maintained cell culture performance.
  • Capacitance measurements are a reliable tool for real-time VCD monitoring and process scale-up in perfusion bioreactors.