Departmental Papers (Dental)
Document Type
Journal Article
Date of this Version
3-2003
Publication Source
Plant Biotechnology Journal
Volume
1
Issue
2
Start Page
71
Last Page
79
DOI
10.1046/j.1467-7652.2003.00008.x
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.
Copyright/Permission Statement
This is the peer reviewed version of the following article: Millán, A. F.-S., Mingo-Castel, A., Miller, M., & Daniell, H. (2003). A chloroplast transgenic approach to hyper-express and purify Human Serum Albumin, a protein highly susceptible to proteolytic degradation. Plant Biotechnology Journal, 1(2), 71–79. http://doi.org/10.1046/j.1467-7652.2003.00008.x, which has been published in final form at 10.1046/j.1467-7652.2003.00008.x. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving http://olabout.wiley.com/WileyCDA/Section/id-820227.html#terms.
Keywords
chloroplast genetic engineering, biopharmaceuticals, genetically modified crops, molecular farming, recombinant human blood proteins
Recommended Citation
Millán, A. F., Mingo-Castel, A., Miller, M., & Daniell, H. (2003). A Chloroplast Transgenic Approach to Hyper-Express and Purify Human Serum Albumin, a Protein Highly Susceptible to Proteolytic Degradation. Plant Biotechnology Journal, 1 (2), 71-79. http://dx.doi.org/10.1046/j.1467-7652.2003.00008.x
Date Posted: 01 March 2022
This document has been peer reviewed.
Comments
At the time of publication, author Henry Daniell was affiliated with the University of Central Florida. Currently, he is a faculty member at the School of Dental Medicine at the University of Pennsylvania.