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State-of-the-art and novel approaches to mild solubilization of inclusion bodies

Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant proteins. Inclusion bodies can easily be separated from the crude extract after cell lysis and contain the product in high purity. How...

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Autores principales: Klausser, Robert, Kopp, Julian, Prada Brichtova, Eva, Gisperg, Florian, Elshazly, Mohamed, Spadiut, Oliver
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10399460/
https://www.ncbi.nlm.nih.gov/pubmed/37545893
http://dx.doi.org/10.3389/fbioe.2023.1249196
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author Klausser, Robert
Kopp, Julian
Prada Brichtova, Eva
Gisperg, Florian
Elshazly, Mohamed
Spadiut, Oliver
author_facet Klausser, Robert
Kopp, Julian
Prada Brichtova, Eva
Gisperg, Florian
Elshazly, Mohamed
Spadiut, Oliver
author_sort Klausser, Robert
collection PubMed
description Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant proteins. Inclusion bodies can easily be separated from the crude extract after cell lysis and contain the product in high purity. However, additional solubilization and refolding steps are required in the processing of IBs to recover the native protein. These unit operations remain a highly empirical field of research in which processes are developed on a case-by-case basis using elaborate screening strategies. It has been shown that a reduction in denaturant concentration during protein solubilization can increase the subsequent refolding yield due to the preservation of correctly folded protein structures. Therefore, many novel solubilization techniques have been developed in the pursuit of mild solubilization conditions that avoid total protein denaturation. In this respect, ionic liquids have been investigated as promising agents, being able to solubilize amyloid-like aggregates and stabilize correctly folded protein structures at the same time. This review briefly summarizes the state-of-the-art of mild solubilization of IBs and highlights some challenges that prevent these novel techniques from being yet adopted in industry. We suggest mechanistic models based on the thermodynamics of protein unfolding with the aid of molecular dynamics simulations as a possible approach to solve these challenges in the future.
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spelling pubmed-103994602023-08-04 State-of-the-art and novel approaches to mild solubilization of inclusion bodies Klausser, Robert Kopp, Julian Prada Brichtova, Eva Gisperg, Florian Elshazly, Mohamed Spadiut, Oliver Front Bioeng Biotechnol Bioengineering and Biotechnology Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant proteins. Inclusion bodies can easily be separated from the crude extract after cell lysis and contain the product in high purity. However, additional solubilization and refolding steps are required in the processing of IBs to recover the native protein. These unit operations remain a highly empirical field of research in which processes are developed on a case-by-case basis using elaborate screening strategies. It has been shown that a reduction in denaturant concentration during protein solubilization can increase the subsequent refolding yield due to the preservation of correctly folded protein structures. Therefore, many novel solubilization techniques have been developed in the pursuit of mild solubilization conditions that avoid total protein denaturation. In this respect, ionic liquids have been investigated as promising agents, being able to solubilize amyloid-like aggregates and stabilize correctly folded protein structures at the same time. This review briefly summarizes the state-of-the-art of mild solubilization of IBs and highlights some challenges that prevent these novel techniques from being yet adopted in industry. We suggest mechanistic models based on the thermodynamics of protein unfolding with the aid of molecular dynamics simulations as a possible approach to solve these challenges in the future. Frontiers Media S.A. 2023-07-20 /pmc/articles/PMC10399460/ /pubmed/37545893 http://dx.doi.org/10.3389/fbioe.2023.1249196 Text en Copyright © 2023 Klausser, Kopp, Prada Brichtova, Gisperg, Elshazly and Spadiut. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Klausser, Robert
Kopp, Julian
Prada Brichtova, Eva
Gisperg, Florian
Elshazly, Mohamed
Spadiut, Oliver
State-of-the-art and novel approaches to mild solubilization of inclusion bodies
title State-of-the-art and novel approaches to mild solubilization of inclusion bodies
title_full State-of-the-art and novel approaches to mild solubilization of inclusion bodies
title_fullStr State-of-the-art and novel approaches to mild solubilization of inclusion bodies
title_full_unstemmed State-of-the-art and novel approaches to mild solubilization of inclusion bodies
title_short State-of-the-art and novel approaches to mild solubilization of inclusion bodies
title_sort state-of-the-art and novel approaches to mild solubilization of inclusion bodies
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10399460/
https://www.ncbi.nlm.nih.gov/pubmed/37545893
http://dx.doi.org/10.3389/fbioe.2023.1249196
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