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dc.creatorFernández San Millán, Aliciaes_ES
dc.creatorMingo Castel, Ángeles_ES
dc.creatorMiller, Michaeles_ES
dc.creatorDaniell, Henryes_ES
dc.identifier.issn1467-7644 (Print)
dc.identifier.issn1467-7652 (Electronic)
dc.description.abstractHuman 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.sponsorshipThis 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.extent9 p.
dc.publisherWiley / Blackwellen
dc.relation.ispartofPlant Biotechnology Journal, (2003) 1, pp. 71–79en
dc.rights© 2003 Blackwell Publishing Ltd. Creative Commons Attribution 4.0 International (CC BY 4.0)en
dc.subjectChloroplast genetic engineeringen
dc.subjectGenetically modified cropsen
dc.subjectMolecular farmingen
dc.subjectRecombinant human blood proteinsen
dc.titleA chloroplast transgenic approach to hyper-express and purify human serum albumin, a protein highly susceptible to proteolytic degradationen
dc.typeArtículo / Artikuluaes
dc.contributor.departmentIdAB - Instituto de Agrobiotecnología / Agrobioteknologiako Institutuaes
dc.rights.accessRightsAcceso abierto / Sarbide irekiaes
dc.type.versionVersión publicada / Argitaratu den bertsioaes
dc.contributor.funderGobierno de Navarra / Nafarroako Gobernuaes

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© 2003 Blackwell Publishing Ltd. Creative Commons Attribution 4.0 International (CC BY 4.0)
Except where otherwise noted, this item's license is described as © 2003 Blackwell Publishing Ltd. Creative Commons Attribution 4.0 International (CC BY 4.0)