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Rapid recombinant protein expression in cell-free extracts from human blood

Several groups have recently reported on the utility of cell-free expression systems to make therapeutic proteins, most of them employing CHO or E. coli cell-free extracts. Here, we propose an alternative that uses human blood derived leukocyte cell extracts for the expression of recombinant protein...

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Autores principales: Burgenson, David, Gurramkonda, Chandrasekhar, Pilli, Manohar, Ge, Xudong, Andar, Abhay, Kostov, Yordan, Tolosa, Leah, Rao, Govind
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6014972/
https://www.ncbi.nlm.nih.gov/pubmed/29934577
http://dx.doi.org/10.1038/s41598-018-27846-8
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author Burgenson, David
Gurramkonda, Chandrasekhar
Pilli, Manohar
Ge, Xudong
Andar, Abhay
Kostov, Yordan
Tolosa, Leah
Rao, Govind
author_facet Burgenson, David
Gurramkonda, Chandrasekhar
Pilli, Manohar
Ge, Xudong
Andar, Abhay
Kostov, Yordan
Tolosa, Leah
Rao, Govind
author_sort Burgenson, David
collection PubMed
description Several groups have recently reported on the utility of cell-free expression systems to make therapeutic proteins, most of them employing CHO or E. coli cell-free extracts. Here, we propose an alternative that uses human blood derived leukocyte cell extracts for the expression of recombinant proteins. We demonstrate expression of nano luciferase (Nluc), Granulocyte-colony stimulating factor (G-CSF) and Erythropoietin (EPO) in cell-free leukocyte extracts within two hours. Human blood is readily available from donors and blood banks and leukocyte rich fractions are easy to obtain. The method described here demonstrates the ability to rapidly express recombinant proteins from human cell extracts that could provide the research community with a facile technology to make their target protein. Eventually, we envision that any recombinant protein can be produced from patient-supplied leukocytes, which can then be injected back into the patient. This approach could lead to an alternative model for personalized medicines and vaccines.
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spelling pubmed-60149722018-06-27 Rapid recombinant protein expression in cell-free extracts from human blood Burgenson, David Gurramkonda, Chandrasekhar Pilli, Manohar Ge, Xudong Andar, Abhay Kostov, Yordan Tolosa, Leah Rao, Govind Sci Rep Article Several groups have recently reported on the utility of cell-free expression systems to make therapeutic proteins, most of them employing CHO or E. coli cell-free extracts. Here, we propose an alternative that uses human blood derived leukocyte cell extracts for the expression of recombinant proteins. We demonstrate expression of nano luciferase (Nluc), Granulocyte-colony stimulating factor (G-CSF) and Erythropoietin (EPO) in cell-free leukocyte extracts within two hours. Human blood is readily available from donors and blood banks and leukocyte rich fractions are easy to obtain. The method described here demonstrates the ability to rapidly express recombinant proteins from human cell extracts that could provide the research community with a facile technology to make their target protein. Eventually, we envision that any recombinant protein can be produced from patient-supplied leukocytes, which can then be injected back into the patient. This approach could lead to an alternative model for personalized medicines and vaccines. Nature Publishing Group UK 2018-06-22 /pmc/articles/PMC6014972/ /pubmed/29934577 http://dx.doi.org/10.1038/s41598-018-27846-8 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Burgenson, David
Gurramkonda, Chandrasekhar
Pilli, Manohar
Ge, Xudong
Andar, Abhay
Kostov, Yordan
Tolosa, Leah
Rao, Govind
Rapid recombinant protein expression in cell-free extracts from human blood
title Rapid recombinant protein expression in cell-free extracts from human blood
title_full Rapid recombinant protein expression in cell-free extracts from human blood
title_fullStr Rapid recombinant protein expression in cell-free extracts from human blood
title_full_unstemmed Rapid recombinant protein expression in cell-free extracts from human blood
title_short Rapid recombinant protein expression in cell-free extracts from human blood
title_sort rapid recombinant protein expression in cell-free extracts from human blood
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6014972/
https://www.ncbi.nlm.nih.gov/pubmed/29934577
http://dx.doi.org/10.1038/s41598-018-27846-8
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