Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture
Summary: Mitogenic growth factors are major cost drivers in serum-free media, contributing up to 95% of the total cost. Here, we present a streamlined workflow detailing cloning, expression testing, protein purification, and bioactivity screening that allows for low-cost production of bioactive grow...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723003180 |
| _version_ | 1797803840051871744 |
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| author | Cameron Semper Alexei Savchenko |
| author_facet | Cameron Semper Alexei Savchenko |
| author_sort | Cameron Semper |
| collection | DOAJ |
| description | Summary: Mitogenic growth factors are major cost drivers in serum-free media, contributing up to 95% of the total cost. Here, we present a streamlined workflow detailing cloning, expression testing, protein purification, and bioactivity screening that allows for low-cost production of bioactive growth factors including basic fibroblast growth factor and transforming growth factor β1. This generalized procedure can be used for multiple families of growth factors with minor modification, and the outputs are bioactive and suitable for cell culture applications.For complete details on the use and execution of this protocol, please refer to Venkatesan, et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-13T05:27:04Z |
| format | Article |
| id | doaj.art-10cda480ad174967a2be468e9dc41ab1 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-13T05:27:04Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-10cda480ad174967a2be468e9dc41ab12023-06-15T04:57:08ZengElsevierSTAR Protocols2666-16672023-09-0143102351Protein expression and purification of bioactive growth factors for use in cell culture and cellular agricultureCameron Semper0Alexei Savchenko1Department of Microbiology, Immunology and Infectious Disease, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada; Corresponding authorDepartment of Microbiology, Immunology and Infectious Disease, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E8, Canada; Corresponding authorSummary: Mitogenic growth factors are major cost drivers in serum-free media, contributing up to 95% of the total cost. Here, we present a streamlined workflow detailing cloning, expression testing, protein purification, and bioactivity screening that allows for low-cost production of bioactive growth factors including basic fibroblast growth factor and transforming growth factor β1. This generalized procedure can be used for multiple families of growth factors with minor modification, and the outputs are bioactive and suitable for cell culture applications.For complete details on the use and execution of this protocol, please refer to Venkatesan, et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723003180Molecular BiologyProtein BiochemistryTissue Engineering |
| spellingShingle | Cameron Semper Alexei Savchenko Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture STAR Protocols Molecular Biology Protein Biochemistry Tissue Engineering |
| title | Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture |
| title_full | Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture |
| title_fullStr | Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture |
| title_full_unstemmed | Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture |
| title_short | Protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture |
| title_sort | protein expression and purification of bioactive growth factors for use in cell culture and cellular agriculture |
| topic | Molecular Biology Protein Biochemistry Tissue Engineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723003180 |
| work_keys_str_mv | AT cameronsemper proteinexpressionandpurificationofbioactivegrowthfactorsforuseincellcultureandcellularagriculture AT alexeisavchenko proteinexpressionandpurificationofbioactivegrowthfactorsforuseincellcultureandcellularagriculture |