Optimization of multi-column chromatography for capture and polishing at high protein load
Silva, Tiago Castanheira ; Isaksson, Madelène LU ; Nilsson, Bernt LU
; Eppink, Michel
and Ottens, Marcel
(2025)
In Biotechnology Progress
41(5).
- Abstract
Integrated Continuous Biomanufacturing reduces manufacturing costs while maintaining product quality. A key contributor to high biopharmaceutical costs, specifically monoclonal antibodies (mAbs), is chromatography. Protein A ligands are usually preferred but still expensive in the manufacturing context, and batch chromatography under-utilizes the columns' capacity, compromising productivity to maintain high yields. Continuous chromatography increases columns' Capacity Utilization (CU) without sacrificing yield or productivity. This work presents the in-silico optimization of a 3 Column Periodic Counter-current Chromatography (3C-PCC) of a capture and polishing step for mAbs from a high titer harvest (cmAb = 5 g/L). The 3C-PCC... (More)
Integrated Continuous Biomanufacturing reduces manufacturing costs while maintaining product quality. A key contributor to high biopharmaceutical costs, specifically monoclonal antibodies (mAbs), is chromatography. Protein A ligands are usually preferred but still expensive in the manufacturing context, and batch chromatography under-utilizes the columns' capacity, compromising productivity to maintain high yields. Continuous chromatography increases columns' Capacity Utilization (CU) without sacrificing yield or productivity. This work presents the in-silico optimization of a 3 Column Periodic Counter-current Chromatography (3C-PCC) of a capture and polishing step for mAbs from a high titer harvest (cmAb = 5 g/L). The 3C-PCC was modeled and Pareto-fronts for continuous and batch modes were used to optimize the 3C-PCC steps varying the flow rate and percentage of breakthrough achieved in the interconnected loading, maximizing Productivity and CU, for varying concentrations of mAb (batch mode concentration of 5 g/L and continuous mode concentration of 2.5, 5, 7.5, and 10 g/L). The shape of the breakthrough curve significantly impacts the optimization of 3C-PCC. The model output was validated for three different protein A ligands using a pure mAb solution. MAb Select SuRe pcc was selected to continuously capture mAb from a high-titer clarified cell culture supernatant (harvest). The product eluates were pooled and used for continuous polishing using a Cation-Exchange resin (CaptoS ImpAct). Experimental results validated model predictions (<7% deviation in the worst case) and a process with two 3C-PCC in sequence was proposed, with a productivity of approximately 100 mg/mL res/h.
(Less)
- author
- Silva, Tiago Castanheira
; Isaksson, Madelène
LU
; Nilsson, Bernt
LU
; Eppink, Michel
and Ottens, Marcel
- organization
- publishing date
- 2025-09-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- continuous chromatography, high titers, integrated continuous biomanufacturing, modeling, periodic counter-current chromatography
- in
- Biotechnology Progress
- volume
- 41
- issue
- 5
- article number
- e70047
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- pmid:40485304
- scopus:105007938775
- ISSN
- 8756-7938
- DOI
- 10.1002/btpr.70047
- language
- English
- LU publication?
- yes
- additional info
- Publisher Copyright: © 2025 The Author(s). Biotechnology Progress published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.
- id
- ca24d6a4-12e0-449a-9700-1cef8bc9d2c4
- date added to LUP
- 2026-01-15 16:37:53
- date last changed
- 2026-01-29 17:58:30
@article{ca24d6a4-12e0-449a-9700-1cef8bc9d2c4,
abstract = {{<p>Integrated Continuous Biomanufacturing reduces manufacturing costs while maintaining product quality. A key contributor to high biopharmaceutical costs, specifically monoclonal antibodies (mAbs), is chromatography. Protein A ligands are usually preferred but still expensive in the manufacturing context, and batch chromatography under-utilizes the columns' capacity, compromising productivity to maintain high yields. Continuous chromatography increases columns' Capacity Utilization (CU) without sacrificing yield or productivity. This work presents the in-silico optimization of a 3 Column Periodic Counter-current Chromatography (3C-PCC) of a capture and polishing step for mAbs from a high titer harvest (c<sub>mAb</sub> = 5 g/L). The 3C-PCC was modeled and Pareto-fronts for continuous and batch modes were used to optimize the 3C-PCC steps varying the flow rate and percentage of breakthrough achieved in the interconnected loading, maximizing Productivity and CU, for varying concentrations of mAb (batch mode concentration of 5 g/L and continuous mode concentration of 2.5, 5, 7.5, and 10 g/L). The shape of the breakthrough curve significantly impacts the optimization of 3C-PCC. The model output was validated for three different protein A ligands using a pure mAb solution. MAb Select SuRe pcc was selected to continuously capture mAb from a high-titer clarified cell culture supernatant (harvest). The product eluates were pooled and used for continuous polishing using a Cation-Exchange resin (CaptoS ImpAct). Experimental results validated model predictions (<7% deviation in the worst case) and a process with two 3C-PCC in sequence was proposed, with a productivity of approximately 100 mg/mL res/h.</p>}},
author = {{Silva, Tiago Castanheira and Isaksson, Madelène and Nilsson, Bernt and Eppink, Michel and Ottens, Marcel}},
issn = {{8756-7938}},
keywords = {{continuous chromatography; high titers; integrated continuous biomanufacturing; modeling; periodic counter-current chromatography}},
language = {{eng}},
month = {{09}},
number = {{5}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Biotechnology Progress}},
title = {{Optimization of multi-column chromatography for capture and polishing at high protein load}},
url = {{http://dx.doi.org/10.1002/btpr.70047}},
doi = {{10.1002/btpr.70047}},
volume = {{41}},
year = {{2025}},
}