Yeast display: Difference between revisions
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'''Yeast display''' (or '''yeast surface display''') is a technique used in the field of [[protein engineering]]. The yeast display technique was |
'''Yeast display''' (or '''yeast surface display''') is a technique used in the field of [[protein engineering]]. The yeast display technique was first published by the [[laboratory]] of Professor K. Dane Wittrup.<ref>[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=AbstractPlus&list_uids=9181578&query_hl=1&itool=pubmed_docsum First yeast display publication]</ref> The technology was sold to Abbott Laboratories in 2001.<ref>[http://www.news.uiuc.edu/NEWS/01/1221biodisplaytechnology.html http://www.news.uiuc.edu/NEWS/01/1221biodisplaytechnology.html]</ref> |
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A protein of interest is displayed as a fusion to the Aga2p protein on the surface of [[yeast]]. The Aga2p protein is naturally used by yeast to mediate cell-cell contacts during yeast cell mating. As such, display of a protein via Aga2p projects the protein away from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of [[flow cytometry]] in conjunction with a yeast display library is a highly effective method to isolate high affinity [[protein]] [[ligand (biochemistry)|ligand]]s against nearly any [[receptor (biochemistry)|receptor]] through [[directed evolution]]. |
A protein of interest is displayed as a fusion to the Aga2p protein on the surface of [[yeast]]. The Aga2p protein is naturally used by yeast to mediate cell-cell contacts during yeast cell mating. As such, display of a protein via Aga2p projects the protein away from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of [[flow cytometry]] in conjunction with a yeast display library is a highly effective method to isolate high affinity [[protein]] [[ligand (biochemistry)|ligand]]s against nearly any [[receptor (biochemistry)|receptor]] through [[directed evolution]]. |
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* Feldhaus M, Siegel R.; Methods in Molecular Biology 263:311-332 (2004). |
* Feldhaus M, Siegel R.; Methods in Molecular Biology 263:311-332 (2004). |
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* Weaver-Feldhaus JM, Lou J, Coleman JR, Siegel RW, Marks JD, Feldhaus MJ. FEBS Lett. 2004 Apr 23;564(1-2):24-34. PMID: 15094038 |
* Weaver-Feldhaus JM, Lou J, Coleman JR, Siegel RW, Marks JD, Feldhaus MJ. FEBS Lett. 2004 Apr 23;564(1-2):24-34. PMID: 15094038 |
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==Notes== |
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Revision as of 20:33, 1 November 2006
Yeast display (or yeast surface display) is a technique used in the field of protein engineering. The yeast display technique was first published by the laboratory of Professor K. Dane Wittrup.[1] The technology was sold to Abbott Laboratories in 2001.[2]
A protein of interest is displayed as a fusion to the Aga2p protein on the surface of yeast. The Aga2p protein is naturally used by yeast to mediate cell-cell contacts during yeast cell mating. As such, display of a protein via Aga2p projects the protein away from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of flow cytometry in conjunction with a yeast display library is a highly effective method to isolate high affinity protein ligands against nearly any receptor through directed evolution.
Advantages and Disadvantages of Yeast Display
Advantages of yeast display over other in vitro evolution methods include eukaryotic expression and processing, quality control mechanisms of the eukaryotic secretory pathway, minimal avidity effects, and quantitative library screening through fluorescent-activated cell sorting (FACS).
Disadvantages include smaller mutagenic library sizes compared to alternative methods and differential glycosylation in yeast compared to mammalian cells. It should be noted that these disadvantages have not limited the success of yeast display for a number of applications, including engineering the highest monovalent ligand-binding affinity reported to date for an engineered protein (Boder, E.T. 2000)
Competing methods for protein evolution in vitro are phage display, ribosome display, bacterial display, and mRNA display.
Citations
- Boder, E.T., Wittrup, K.D.; Nat. Biotech., 1997, 15, 553-57.
- Boder, E.T., Wittrup, K.D.; Biotechnol. Prog., 1998, 14, 55-62.
- Boder E.T., Midelfort K.S., Wittrup K.D.; Proc Nat Acad Sci, 2000, 97(20):10701-10705.
- Graff, C.P., Chester, K., Begent, R., Wittrup, K.D.; Prot. Eng. Des. Sel., 2004, 17, 293-304.
- Feldhaus M, Siegel R.; Methods in Molecular Biology 263:311-332 (2004).
- Weaver-Feldhaus JM, Lou J, Coleman JR, Siegel RW, Marks JD, Feldhaus MJ. FEBS Lett. 2004 Apr 23;564(1-2):24-34. PMID: 15094038
Notes
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