Purification of a Hydrophobic Elastin-Like Protein Toward Scale-Suitable Production of Biomaterials.
biomaterial production
hydrophobic elastin-like protein (ELP)
process scalability
protein purification
thermoresponsive protein/polymer
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:
2022
2022
Historique:
received:
18
02
2022
accepted:
10
05
2022
entrez:
11
7
2022
pubmed:
12
7
2022
medline:
12
7
2022
Statut:
epublish
Résumé
Elastin-like proteins (ELPs) are polypeptides with potential applications as renewable bio-based high-performance polymers, which undergo a stimulus-responsive reversible phase transition. The ELP investigated in this manuscript-ELP[V2Y-45]-promises fascinating mechanical properties in biomaterial applications. Purification process scalability and purification performance are important factors for the evaluation of potential industrial-scale production of ELPs. Salt-induced precipitation, inverse transition cycling (ITC), and immobilized metal ion affinity chromatography (IMAC) were assessed as purification protocols for a polyhistidine-tagged hydrophobic ELP showing low-temperature transition behavior. IMAC achieved a purity of 86% and the lowest nucleic acid contamination of all processes. Metal ion leakage did not propagate chemical modifications and could be successfully removed through size-exclusion chromatography. The simplest approach using a high-salt precipitation resulted in a 60% higher target molecule yield compared to both other approaches, with the drawback of a lower purity of 60% and higher nucleic acid contamination. An additional ITC purification led to the highest purity of 88% and high nucleic acid removal. However, expensive temperature-dependent centrifugation steps are required and aggregation effects even at low temperatures have to be considered for the investigated ELP. Therefore, ITC and IMAC are promising downstream processes for biomedical applications with scale-dependent economical costs to be considered, while salt-induced precipitation may be a fast and simple alternative for large-scale bio-based polymer production.
Identifiants
pubmed: 35814018
doi: 10.3389/fbioe.2022.878838
pii: 878838
pmc: PMC9257828
doi:
Types de publication
Journal Article
Langues
eng
Pagination
878838Informations de copyright
Copyright © 2022 Haas, Desombre, Kirschhöfer, Huber, Schiller 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.
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