A chloroplast transgenic approach to hyper-express and purify human serum albumin, a protein highly susceptible to proteolytic degradation
| dc.contributor.author | Fernández San Millán, Alicia | |
| dc.contributor.author | Mingo Castel, Ángel | |
| dc.contributor.author | Miller, Michael | |
| dc.contributor.author | Daniell, Henry | |
| dc.contributor.department | IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua | es_ES |
| dc.contributor.funder | Gobierno de Navarra / Nafarroako Gobernua | es |
| dc.date.accessioned | 2018-11-27T11:22:22Z | |
| dc.date.available | 2018-11-27T11:22:22Z | |
| dc.date.issued | 2003 | |
| dc.description.abstract | Human Serum Albumin (HSA) accounts for 60% of the total protein in blood serum and it is the most widely used intravenous protein in a number of human therapies. HSA, however, is currently extracted only from blood because of a lack of commercially feasible recombinant expression systems. HSA is highly susceptible to proteolytic degradation in recombinant systems and is expensive to purify. Expression of HSA in transgenic chloroplasts using Shine-Dalgarno sequence (SD), which usually facilitates hyper-expression of transgenes, resulted only in 0.02% HSA in total protein (tp). Modification of HSA regulatory sequences using chloroplast untranslated regions (UTRs) resulted in hyper-expression of HSA (up to 11.1% tp), compensating for excessive proteolytic degradation. This is the highest expression of a pharmaceutical protein in transgenic plants and 500-fold greater than previous reports on HSA expression in transgenic leaves. Electron micrographs of immunogold labelled transgenic chloroplasts revealed HSA inclusion bodies, which provided a simple method for purification from other cellular proteins. HSA inclusion bodies could be readily solubilized to obtain a monomeric form using appropriate reagents. The regulatory elements used in this study should serve as a model system for enhancing expression of foreign proteins that are highly susceptible to proteolytic degradation and provide advantages in purification, when inclusion bodies are formed. | en |
| dc.description.sponsorship | This study was supported in part by grants from NIH RO1 GM 63879 and Chlorgen Inc. to H.D., and ‘Dirección General de Industria, Gobierno de Navarra’ (Spain) to A.M.C. | en |
| dc.format.extent | 9 p. | |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.doi | 10.1046/j.1467-7652.2003.00008.x | |
| dc.identifier.issn | 1467-7644 (Print) | |
| dc.identifier.issn | 1467-7652 (Electronic) | |
| dc.identifier.uri | https://academica-e.unavarra.es/handle/2454/31507 | |
| dc.language.iso | eng | en |
| dc.publisher | Wiley / Blackwell | en |
| dc.relation.ispartof | Plant Biotechnology Journal, (2003) 1, pp. 71–79 | en |
| dc.relation.publisherversion | https://doi.org/10.1046/j.1467-7652.2003.00008.x | |
| dc.rights | © 2003 Blackwell Publishing Ltd. Creative Commons Attribution 4.0 International (CC BY 4.0) | en |
| dc.rights.accessRights | info:eu-repo/semantics/openAccess | |
| dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
| dc.subject | Chloroplast genetic engineering | en |
| dc.subject | Biopharmaceuticals | en |
| dc.subject | Genetically modified crops | en |
| dc.subject | Molecular farming | en |
| dc.subject | Recombinant human blood proteins | en |
| dc.title | A chloroplast transgenic approach to hyper-express and purify human serum albumin, a protein highly susceptible to proteolytic degradation | en |
| dc.type | info:eu-repo/semantics/article | |
| dc.type.version | info:eu-repo/semantics/publishedVersion | |
| dspace.entity.type | Publication | |
| relation.isAuthorOfPublication | 9c84c660-8e77-4487-bd7a-07ad058b0425 | |
| relation.isAuthorOfPublication | 2253e7e5-b778-474f-b91f-d13c494268c1 | |
| relation.isAuthorOfPublication.latestForDiscovery | 9c84c660-8e77-4487-bd7a-07ad058b0425 |