Application of a fluorescent dye-based microfluidic sensor for real-time detection of mAb aggregates.

Autor: São Pedro MN; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands., Eppink MHM; Byondis B.V., Nijmegen, The Netherlands.; Bioprocessing Engineering, Wageningen University, Wageningen, The Netherlands., Ottens M; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.
Jazyk: angličtina
Zdroj: Biotechnology progress [Biotechnol Prog] 2024 Sep-Oct; Vol. 40 (5), pp. e3355. Date of Electronic Publication: 2023 May 10.
DOI: 10.1002/btpr.3355
Abstrakt: The lack of process analytical technologies able to provide real-time information and process control over a biopharmaceutical process has long impaired the transition to continuous biomanufacturing. For the monoclonal antibody (mAb) production, aggregate formation is a major critical quality attribute (CQA) with several known process parameters (i.e., protein concentration and agitation) influencing this phenomenon. The development of a real-time tool to monitor aggregate formation is then crucial to gain control and achieve a continuous processing. Due to an inherent short operation time, miniaturized biosensors placed after each step can be a powerful solution. In this work, the development of a fluorescent dye-based microfluidic sensor for fast at-line PAT is described, using fluorescent dyes to examine possible mAb size differences. A zigzag microchannel, which provides 90% of mixing efficiency under 30 s, coupled to an UV-Vis detector, and using four FDs, was studied and validated. With different generated mAb aggregation samples, the FDs Bis-ANS and CCVJ were able to robustly detect from, at least, 2.5% to 10% of aggregation. The proposed FD-based micromixer is then ultimately implemented and validated in a lab-scale purification system, demonstrating the potential of a miniaturized biosensor to speed up CQAs measurement in a continuous process.
(© 2023 The Authors. Biotechnology Progress published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)
Databáze: MEDLINE
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