Yeast display
Yeast display (or yeast surface display) is a protein engineering technique that uses the expression of recombinant proteins incorporated into the cell wall of yeast for isolating and engineering antibodies.[1]
Development[]
The yeast display technique was first published by the laboratory of Professor K. Dane Wittrup.[2] The technology was sold to Abbott Laboratories in 2001.[3]
How it works[]
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 magnetic separation and 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[]
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). Yeast are eukaryotic organisms that allow for complex post-translational modifications to proteins that no other display libraries are able to provide.
Disadvantages include smaller mutant library sizes compared to alternative methods and differential glycosylation in yeast compared to mammalian cells. Alternative methods for protein evolution in vitro are , phage display, ribosome display, bacterial display, and mRNA display.
References[]
- ^ Gai, S Annie; Wittrup, K Dane (2007). "Yeast surface display for protein engineering and characterization". Current Opinion in Structural Biology. 17 (4): 467–473. doi:10.1016/j.sbi.2007.08.012. ISSN 0959-440X. PMC 4038029. PMID 17870469.
- ^ Boder, Eric T.; Wittrup, K. Dane (1997). "Yeast surface display for screening combinatorial polypeptide libraries". Nature Biotechnology. 15 (6): 553–557. doi:10.1038/nbt0697-553. ISSN 1087-0156. PMID 9181578. S2CID 23922281.
- ^ http://www.news.uiuc.edu/NEWS/01/1221biodisplaytechnology.html
Further reading[]
- Boder, E.T., Wittrup, K.D.; Biotechnol. Prog., 1998, 14, 55–62.
- Boder E.T., Midelfort K.S., Wittrup K.D.; Proc Natl 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, Jane M; Lou, Jianlong; Coleman, James R; Siegel, Robert W; Marks, James D; Feldhaus, Michael J (2004). "Yeast mating for combinatorial Fab library generation and surface display". FEBS Letters. 564 (1–2): 24–34. doi:10.1016/S0014-5793(04)00309-6. ISSN 0014-5793. PMID 15094038. S2CID 29737912.
- Biochemistry methods