Continuous Antibody Titer Assessment of the Biomanufacturing Process using Nanofluidic Binding Assays

Abstract

Online monitoring of monoclonal antibody (mAb) product titers throughout biologics process development and production enables the implementation of quality-by-design biomanufacturing, rapid bioprocess decision-making, and process optimization. Intermittent sampling and analysis are presently utilized to maintain long-term perfusion culture. However, this increases the risk of introducing external contaminants into the cell culture and the analyzed analytes. Sensors used in in-line probe-based approaches must withstand harsh clean-in-place and sterilization-in-place procedures. Analytical instruments that provide real-time monitoring and continuous feed flow processing and are therefore suitable for direct integration into a perfusion bioreactor are ideal for improving overall process robustness. Online implementation of conventional analytical methods, including high-performance liquid chromatography (HPLC) and turbidimetric analysis, typically vii necessitates interfacing with an automated sampling system capable of online sampling and fractionation, which appends to the cost, risk of failure, and mechanical complexity of the system.

The objective of this study is to create a nanofluidic device for online monitoring of monoclonal antibody (mAb) titers based on ligand-binding. This system has a small footprint, straightforward operation procedures, and minimal complex data analytics requirements. During manufacturing and process development, this nanofluidic platform delivers direct titer measurement in the bioreactor as well as functional information on the binding activity, enabling immediate modification of process parameters to improve biomanufacturing yield and preserve the desired product quality.