Impact of freeze-thaw processes on monoclonal antibody platform process development.
cell culture supernatant
freezing
monoclonal antibody
platform process
process development
Journal
Biotechnology and bioengineering
ISSN: 1097-0290
Titre abrégé: Biotechnol Bioeng
Pays: United States
ID NLM: 7502021
Informations de publication
Date de publication:
10 2021
10 2021
Historique:
revised:
16
06
2021
received:
16
04
2021
accepted:
19
06
2021
pubmed:
26
6
2021
medline:
3
3
2022
entrez:
25
6
2021
Statut:
ppublish
Résumé
Freezing of cell culture supernatant (CCS) is a standard procedure in process development of monoclonal antibody (mAb) platform processes as up- and downstream development are usually separated. In the manufacturing process of mAb, however, freezing is avoided, which poses the question of comparability and transferability from process development to manufacturing. In this case study, mAb CCS from Chinese hamster ovary (CHO) cells is frozen and thawed in a novel active freezing device and subsequently captured by protein A chromatography. Critical quality attributes such as host cell protein (HCP) concentration and soluble mAb dimer shares have been monitored throughout the case study. Furthermore, cryo-concentration of individual proteins was investigated. The main factors that drive cryo-concentration are diffusion and natural convection. Natural convection in freezing processes was found to increase at warmer freezing temperatures and thus slower freezing, leading to higher concentration gradients from top to bottom of a freezing chamber. The freeze concentration was dependent on protein size and correlated to diffusivity, where smaller proteins are exposed to higher cryo-concentration. Our results suggest that as a result of freezing processes, large particles based on mAb and specific host cell proteins (HCPs) expressing a certain affinity to mAbs are formed that have to be removed before purification. This leads to a significant improvement in HCP reduction by the protein A step, when compared with reference samples, where twice as much HCP remained in the eluate. Furthermore, HCP and mAb dimer concentrations in protein A eluate were dependent on the freezing temperature. As a conclusion, CCS should be frozen as rapidly as possible during process development to minimize issues of transferability from process development to manufacturing.
Substances chimiques
Antibodies, Monoclonal
0
Culture Media
0
Staphylococcal Protein A
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3914-3925Informations de copyright
© 2021 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.
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