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

São Pedro, Mariana N. ; Isaksson, Madelène LU ; Gomis-Fons, Joaquín LU ; Eppink, Michel H.M. ; Nilsson, Bernt LU orcid and Ottens, Marcel (2023) In Biotechnology and Bioengineering 120(10). p.2989-3000
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... (More)

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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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
antibody aggregation, continuous biomanufacturing, fluorescent dyes, microfluidic sensor, process analytical technology (PAT)
in
Biotechnology and Bioengineering
volume
120
issue
10
pages
12 pages
publisher
John Wiley & Sons Inc.
external identifiers
  • pmid:37309984
  • scopus:85163112413
ISSN
0006-3592
DOI
10.1002/bit.28466
language
English
LU publication?
yes
id
6dbf6261-f030-4c8a-88d2-a47a95b70d32
date added to LUP
2023-11-09 15:06:05
date last changed
2025-03-10 12:56:10
@article{6dbf6261-f030-4c8a-88d2-a47a95b70d32,
  abstract     = {{<p>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.</p>}},
  author       = {{São Pedro, Mariana N. and Isaksson, Madelène and Gomis-Fons, Joaquín and Eppink, Michel H.M. and Nilsson, Bernt and Ottens, Marcel}},
  issn         = {{0006-3592}},
  keywords     = {{antibody aggregation; continuous biomanufacturing; fluorescent dyes; microfluidic sensor; process analytical technology (PAT)}},
  language     = {{eng}},
  number       = {{10}},
  pages        = {{2989--3000}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Biotechnology and Bioengineering}},
  title        = {{Real-time detection of mAb aggregates in an integrated downstream process}},
  url          = {{http://dx.doi.org/10.1002/bit.28466}},
  doi          = {{10.1002/bit.28466}},
  volume       = {{120}},
  year         = {{2023}},
}