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Real‐Time Detection of mAb Aggregates in an Integrated Downstream Process

São Pedro, Mariana N., Isaksson, Madelène, Gomis‐Fons, Joaquín, Eppink, Michel H. M., Nilsson, Bernt, Ottens, Marcel·2023·Biotechnology and Bioengineering
DOI 10.1002/bit.28466
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Abstract

The implementation of continuous processing in the biopharmaceutical industry is hindered by the scarcity of process analytical technologies (PAT). To monitor and control a continuous process, PAT tools will be crucial to measure real‐time product quality attributes such as protein aggregation. Miniaturizing these analytical techniques can increase measurement speed and enable faster decision‐making. A fluorescent dye (FD)‐based miniaturized sensor has previously been developed: a zigzag microchannel which mixes two streams under 30 s. Bis‐ANS and CCVJ, two established FDs, were employed in this micromixer to detect aggregation of the biopharmaceutical monoclonal antibody (mAb). Both FDs were able to robustly detect aggregation levels starting at 2.5%. However, the real‐time measurement provided by the microfluidic sensor still needs to be implemented and assessed in an integrated continuous downstream process. In this work, the micromixer is implemented in a lab‐scale integrated system for the purification of mAbs, established in an ÄKTA™ unit. A viral inactivation and two polishing steps were reproduced, sending a sample of the product pool after each phase directly to the microfluidic sensor for aggregate detection. An additional UV sensor was connected after the micromixer and an increase in its signal would indicate that aggregates were present in the sample. The at‐line miniaturized PAT tool provides a fast aggregation measurement, under 10 min, enabling better process understanding and control.

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  • Pedro et al. miniaturized a fluorescence dye-based microfluidic sensor and measured protein aggregation in under 10 min.

    “Pedro et al., explored this potential by miniaturizing a fluorescence dye–based microfluidic sensor, and the measurement of protein aggregation was performed in under 10 min.”

    From Current PAT Landscape in the Downstream Processing of Biopharmaceuticals (2025) · cites this paper as [94]

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Authors
São Pedro, M. N., Isaksson, M., Gomis‐Fons, J., Eppink, M. H. M., Nilsson, B., & Ottens, M.
Affiliations
  • Department of Biotechnology Delft University of Technology Delft The Netherlands
  • Department of Chemical Engineering Lund University Lund Sweden
  • Byondis B. V. Nijmegen The Netherlands
ORCID
Venue
Biotechnology and Bioengineering
Publisher
Wiley
Published
2023-06-13
Volume
120
Issue
10
Pages
2989-3000
ISSN
  • 0006-3592
  • 1097-0290
DOI
10.1002/bit.28466
Cited by
15
References
1
Language
en
Licence
creativecommons.org/licenses/by-nc-nd/4.0/ (vor)

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