Abstract
Intein-based bioseparations have become a widely used technique for the purification of single proteins at the research scale. Aspects of these purification methods suggest that they might eventually be used in more advanced applications, including large-scale protein production and high-throughput proteomic studies. Both of these applications require substantial analysis for further development, particularly concerning the economics and practicality of potential future systems. For scale-up to commercial protein production, an analysis has been made using widely accepted assumptions for process design and a computer model of a hypothetical process. It is clear that the intein process has the potential to be economically competitive, but will be more attractive with the use of pH- and temperature-controlled inteins with lowcost buffer systems. In the case of proteomic applications, prototypical devices have been constructed to demonstrate feasibility. Both of these studies indicate a strong potential for inteins to eventually join mainstream technologies in these areas.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Chong S, Mersha FB, Comb DG, Scott ME, Landry D, Vence LM, Perler FB, Benner J, Kucera RB, Hirvonen CA, Pelletier JJ, Paulus H, Xu M-Q (1997) Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element. Gene 192:271–281
Chong S, Montello GE, Zhang A, Cantor EJ, Liao W, Xu M-Q, Benner J (1998) Utilizing the Cterminal cleavage activity of a protein splicing element to purify recombinant proteins in a single chromatographic step. Nucleic Acids Res 26:5109–5115
De Serres FJ (2002) Worldwide racial and ethnic distribution of α1-antitrypsin deficiency: summary of an analysis of published genetic epidemiologic surveys. Chest 122:1818–1829
Fish NM, Lilly MD (1984) The interactions between fermentation and protein recovery. Bio/Technology 2:623–627
Germino J, Bastia D (1984) Rapid purification of a cloned gene product by genetic fusion and site-specific proteolysis. Proc Natl Acad Sci USA 81:4692–4696
Harrison RJ, Todd P, Rudge SR, Petrides DP (2003) Bioseparations science and engineering. Oxford Univ Press, New York
Kapust RB, Waugh DS (1999) Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused. Protein Sci 8:1668–1674
Ladisch MR (2001) Bioseparations engineering. Wiley, Hoboken, NJ
LaVallie ER, McCoy JM (1995) Gene fusion expression systems in Escherichia coli. Curr Opin Biotechnol 6:501–506
Lesaicherre ML, Lue RY, Chen GY, Zhu Q, Yao SQ (2002) Intein-mediated biotinylation of proteins and its application in a protein microarray. J Am Chem Soc 124:8768–8769
Lue RY, Chen GY, Hu Y, Zhu Q, Yao SQ (2004) Versatile protein biotinylation strategies for potential high-throughput proteomics. J Am Chem Soc 126:1055–1062
Ma J, Cooney CL (2004) Application of vortex flow adsorption technology to intein-mediated recovery of recombinant human alpha1-antitrypsin. Biotechnol Prog 20:269–276
Miao J, Wu W, Spielmann T, Belfort M, Derbyshire V, Belfort G (2005) Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform. Lab Chip 5:248–253.
Petrides DP, Koulouris A, Lagonikos PT (2002) The role of process simulation in pharmaceutical process development and product commercialization. Pharmaceut Eng 22:56–65
Rouf SA, Douglas PL, Moo-Young M, Scharer JM (2001) Computer simulation for large scale bioprocess design. Biochem Eng J 8:229–234
Shanklin T, Roper K, Yegneswaran PK, Marten MR (2001) Selection of bioprocess simulation software for industrial applications. Biotechnol Bioeng 72:483–489
Sharma S, Zhang A, Wang H, Harcum SW, Chong S (2003) Study of protein splicing and intein-mediated peptide bond cleavage under high-cell-density conditions. Biotechnol Prog 19:1085–1090
Southworth MW, Amaya K, Evans TC, Xu MQ, Perler FB (1999) Purification of proteins fused to either the amino or carboxy terminus of the Mycobacterium xenopi gyrase A intein. Biotechniques 27:110–114, 116, 118–120
Swartz JR (2001) Advances in Escherichia coli production of therapeutic proteins. Curr Opin Biotechnol 12:195–201
Wood DW, Wu W, Belfort G, Derbyshire V, Belfort M (1999) A genetic system yields selfcleaving inteins for bioseparations. Nat Biotechnol 17:889–892
Wood DW, Derbyshire V, Wu W, Chartrain M, Belfort M, Belfort G (2000) Optimized singlestep affinity purification with a self-cleaving intein applied to human acidic fibroblast growth factor. Biotechnol Prog 16:1055–1063
Wu W, Wood DW, Belfort G, Derbyshire V, Belfort M (2002) Intein-mediated purification of cytotoxic endonuclease I-TevI by insertional inactivation and pH-controllable splicing. Nucleic Acids Res 30:4864–4871
Xu MQ, Paulus H, Chong S (2000) Fusions to self-splicing inteins for protein purification. Methods Enzymol 326:376–418
Zhang A, Gonzalez SM, Cantor EJ, Chong S (2001) Construction of a mini-intein fusion system to allow both direct monitoring of soluble protein expression and rapid purification of target proteins. Gene 275:241–252
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Wood, D.W., Harcum, S.W., Belfort, G. (2005). Industrial Applications of Intein Technology. In: Belfort, M., Wood, D.W., Stoddard, B.L., Derbyshire, V. (eds) Homing Endonucleases and Inteins. Nucleic Acids and Molecular Biology, vol 16. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-29474-0_20
Download citation
DOI: https://doi.org/10.1007/3-540-29474-0_20
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-25106-4
Online ISBN: 978-3-540-29474-0
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)