Real-time detection of mAb aggregates in an integrated downstream process

Journal Article (2023)
Author(s)

Mariana N. Neves Sao Pedro (TU Delft - BT/Bioprocess Engineering)

Madelène Isaksson (Lund University)

Joaquín Gomis-Fons (Lund University)

Michel H.M. Eppink (Wageningen University & Research, Byondis B.V., Nijmegen)

Bernt Nilsson (Lund University)

M Ottens (TU Delft - BT/Design and Engineering Education)

Research Group
BT/Design and Engineering Education
Copyright
© 2023 M. Neves Sao Pedro, Madelène Isaksson, Joaquín Gomis-Fons, Michel H.M. Eppink, Bernt Nilsson, M. Ottens
DOI related publication
https://doi.org/10.1002/bit.28466
More Info
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Publication Year
2023
Language
English
Copyright
© 2023 M. Neves Sao Pedro, Madelène Isaksson, Joaquín Gomis-Fons, Michel H.M. Eppink, Bernt Nilsson, M. Ottens
Related content
Research Group
BT/Design and Engineering Education
Issue number
10
Volume number
120
Pages (from-to)
2989-3000
Reuse Rights

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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.