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Design of an integrated continuous downstream process for acid-sensitive monoclonal antibodies based on a calcium-dependent Protein A ligand

Scheffel, Julia ; Isaksson, Madelène LU ; Gomis-Fons, Joaquín LU ; Schwarz, Hubert ; Andersson, Niklas LU orcid ; Norén, Björn ; Solbrand, Anita ; Chotteau, Veronique ; Hober, Sophia and Nilsson, Bernt LU (2022) In Journal of Chromatography A 1664.
Abstract
Monoclonal antibodies (mAb) are used as therapeutics and for diagnostics of a variety of diseases, and novel antibodies are continuously being developed to find treatments for new diseases. Therefore, the manufacturing process must accommodate a range of mAb characteristics. Acid-sensitive mAbs can severely compromise product purity and yield in the purification process due to the potential formation of aggregates. To address this problem, we have developed an integrated downstream process for the purification of pH-sensitive mAbs at mild conditions. A calcium-dependent Protein A-based ligand, called ZCa, was used in the capture step in a 3-column periodic counter-current chromatography operation. The binding of ZCa... (More)
Monoclonal antibodies (mAb) are used as therapeutics and for diagnostics of a variety of diseases, and novel antibodies are continuously being developed to find treatments for new diseases. Therefore, the manufacturing process must accommodate a range of mAb characteristics. Acid-sensitive mAbs can severely compromise product purity and yield in the purification process due to the potential formation of aggregates. To address this problem, we have developed an integrated downstream process for the purification of pH-sensitive mAbs at mild conditions. A calcium-dependent Protein A-based ligand, called ZCa, was used in the capture step in a 3-column periodic counter-current chromatography operation. The binding of ZCa to antibodies is regulated by calcium, meaning that acidic conditions are not needed to break the interaction and elute the antibodies. Further, the virus inactivation was achieved by a solvent/detergent method, where the pH could remain unchanged. The polishing steps included a cation and an anion exchange chromatography step, and screening of the capture and polishing steps was performed to allow for a seamless integration of the process steps. The process was implemented at laboratory scale for 9 days obtaining a high yield, and a consistently pure drug substance, including high reduction values of the host cell protein and DNA concentrations, as well as aggregate levels below the detection limit, which is attributed to the mild conditions used in the process. (Less)
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author
; ; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Monoclonal antibody, Integrated continuous downstream process, pH-sensitive antibodies, Mild purification, ZCa, Process design
in
Journal of Chromatography A
volume
1664
article number
462806
pages
8 pages
publisher
Elsevier
external identifiers
  • pmid:35033788
  • scopus:85122694908
ISSN
0021-9673
DOI
10.1016/j.chroma.2022.462806
language
English
LU publication?
yes
id
94eb629d-45db-4a07-8c13-0a06dc778f3a
date added to LUP
2022-02-03 13:11:26
date last changed
2023-12-11 14:49:35
@article{94eb629d-45db-4a07-8c13-0a06dc778f3a,
  abstract     = {{Monoclonal antibodies (mAb) are used as therapeutics and for diagnostics of a variety of diseases, and novel antibodies are continuously being developed to find treatments for new diseases. Therefore, the manufacturing process must accommodate a range of mAb characteristics. Acid-sensitive mAbs can severely compromise product purity and yield in the purification process due to the potential formation of aggregates. To address this problem, we have developed an integrated downstream process for the purification of pH-sensitive mAbs at mild conditions. A calcium-dependent Protein A-based ligand, called Z<sub>Ca</sub>, was used in the capture step in a 3-column periodic counter-current chromatography operation. The binding of Z<sub>Ca</sub> to antibodies is regulated by calcium, meaning that acidic conditions are not needed to break the interaction and elute the antibodies. Further, the virus inactivation was achieved by a solvent/detergent method, where the pH could remain unchanged. The polishing steps included a cation and an anion exchange chromatography step, and screening of the capture and polishing steps was performed to allow for a seamless integration of the process steps. The process was implemented at laboratory scale for 9 days obtaining a high yield, and a consistently pure drug substance, including high reduction values of the host cell protein and DNA concentrations, as well as aggregate levels below the detection limit, which is attributed to the mild conditions used in the process.}},
  author       = {{Scheffel, Julia and Isaksson, Madelène and Gomis-Fons, Joaquín and Schwarz, Hubert and Andersson, Niklas and Norén, Björn and Solbrand, Anita and Chotteau, Veronique and Hober, Sophia and Nilsson, Bernt}},
  issn         = {{0021-9673}},
  keywords     = {{Monoclonal antibody; Integrated continuous downstream process; pH-sensitive antibodies; Mild purification; ZCa; Process design}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Chromatography A}},
  title        = {{Design of an integrated continuous downstream process for acid-sensitive monoclonal antibodies based on a calcium-dependent Protein A ligand}},
  url          = {{http://dx.doi.org/10.1016/j.chroma.2022.462806}},
  doi          = {{10.1016/j.chroma.2022.462806}},
  volume       = {{1664}},
  year         = {{2022}},
}