Spectroscopic insights into multi-phase protein crystallization in complex lysate using Raman spectroscopy and a particle-free bypass.

E. coli lysate Raman spectroscopy capture process partial least squares regression (PLS) principal component analyses (PCA) process analytical technology (PAT) protein crystallization ultraviolet-visible light (UV/Vis) spectroscopy

Journal

Frontiers in bioengineering and biotechnology
ISSN: 2296-4185
Titre abrégé: Front Bioeng Biotechnol
Pays: Switzerland
ID NLM: 101632513

Informations de publication

Date de publication:
2024
Historique:
received: 07 03 2024
accepted: 23 04 2024
medline: 30 5 2024
pubmed: 30 5 2024
entrez: 30 5 2024
Statut: epublish

Résumé

Protein crystallization as opposed to well-established chromatography processes has the benefits to reduce production costs while reaching a comparable high purity. However, monitoring crystallization processes remains a challenge as the produced crystals may interfere with analytical measurements. Especially for capturing proteins from complex feedstock containing various impurities, establishing reliable process analytical technology (PAT) to monitor protein crystallization processes can be complicated. In heterogeneous mixtures, important product characteristics can be found by multivariate analysis and chemometrics, thus contributing to the development of a thorough process understanding. In this project, an analytical set-up is established combining offline analytics, on-line ultraviolet visible light (UV/Vis) spectroscopy, and in-line Raman spectroscopy to monitor a stirred-batch crystallization process with multiple phases and species being present. As an example process, the enzyme

Identifiants

pubmed: 38812919
doi: 10.3389/fbioe.2024.1397465
pii: 1397465
pmc: PMC11133712
doi:

Types de publication

Journal Article

Langues

eng

Pagination

1397465

Informations de copyright

Copyright © 2024 Wegner, Eming, Walla, Bischoff, Weuster-Botz and Hubbuch.

Déclaration de conflit d'intérêts

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Auteurs

Christina Henriette Wegner (CH)

Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.

Sebastian Mathis Eming (SM)

Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.

Brigitte Walla (B)

Institute of Biochemical Engineering, Technical University of Munich, Garching, Germany.

Daniel Bischoff (D)

Institute of Biochemical Engineering, Technical University of Munich, Garching, Germany.

Dirk Weuster-Botz (D)

Institute of Biochemical Engineering, Technical University of Munich, Garching, Germany.

Jürgen Hubbuch (J)

Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.

Classifications MeSH