Yeast display (or yeast surface display) is a
protein engineering
Protein engineering is the process of developing useful or valuable proteins through the design and production of unnatural polypeptides, often by altering amino acid sequences found in nature. It is a young discipline, with much research taking pl ...
technique that uses the expression of
recombinant protein
Protein production is the biotechnological process of generating a specific protein. It is typically achieved by the manipulation of gene expression in an organism such that it expresses large amounts of a recombinant gene. This includes the ...
s incorporated into the cell wall of
yeast
Yeasts are eukaryotic, single-celled microorganisms classified as members of the fungus kingdom (biology), kingdom. The first yeast originated hundreds of millions of years ago, and at least 1,500 species are currently recognized. They are est ...
. This method can be used for several applications such as isolating and engineering
antibodies
An antibody (Ab) or immunoglobulin (Ig) is a large, Y-shaped protein belonging to the immunoglobulin superfamily which is used by the immune system to identify and neutralize antigens such as bacteria and viruses, including those that caus ...
and determining host-microbe interactions.
Development
The yeast display technique was first published by the
laboratory
A laboratory (; ; colloquially lab) is a facility that provides controlled conditions in which scientific or technological research, experiments, and measurement may be performed. Laboratories are found in a variety of settings such as schools ...
of Professor K. Dane Wittrup and Eric T. Boder.
The technology was sold to
Abbott Laboratories
Abbott Laboratories is an American multinational medical devices and health care company with headquarters in Abbott Park, Illinois, in the United States. The company was founded by Chicago physician Wallace Calvin Abbott in 1888 to formulate k ...
in 2001.
http://www.news.uiuc.edu/NEWS/01/1221biodisplaytechnology.html
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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 used by yeast to mediate cell–cell contacts during yeast cell mating. As such, display of a protein via Aga2p likely projects the fusion protein from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of magnetic separation
Magnetic separation is the process of separating components of mixtures by using a magnet to attract magnetic substances. The process that is used for magnetic separation separates non-magnetic substances from those which are magnetic. This techniq ...
and flow cytometry
Flow cytometry (FC) is a technique used to detect and measure the physical and chemical characteristics of a population of cells or particles.
In this process, a sample containing cells or particles is suspended in a fluid and injected into the ...
in conjunction with a yeast display library can be highly effective method to isolate high affinity protein
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residue (biochemistry), residues. Proteins perform a vast array of functions within organisms, including Enzyme catalysis, catalysing metab ...
ligand
In coordination chemistry, a ligand is an ion or molecule with a functional group that binds to a central metal atom to form a coordination complex. The bonding with the metal generally involves formal donation of one or more of the ligand's el ...
s against nearly any receptor
Receptor may refer to:
* Sensory receptor, in physiology, any neurite structure that, on receiving environmental stimuli, produces an informative nerve impulse
*Receptor (biochemistry), in biochemistry, a protein molecule that receives and respond ...
through directed evolution
Directed evolution (DE) is a method used in protein engineering that mimics the process of natural selection to steer proteins or nucleic acids toward a user-defined goal. It consists of subjecting a gene to iterative rounds of mutagenesis (cre ...
.
Advantages and disadvantages
Advantages of yeast display over other ''in vitro'' evolution methods include eukaryotic
The eukaryotes ( ) constitute the Domain (biology), domain of Eukaryota or Eukarya, organisms whose Cell (biology), cells have a membrane-bound cell nucleus, nucleus. All animals, plants, Fungus, fungi, seaweeds, and many unicellular organisms ...
expression and post translational 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
Glycosylation is the reaction in which a carbohydrate (or ' glycan'), i.e. a glycosyl donor, is attached to a hydroxyl or other functional group of another molecule (a glycosyl acceptor) in order to form a glycoconjugate. In biology (but not ...
in yeast compared to mammalian cells. Alternative methods for protein evolution ''in vitro'' are mammalian display, phage display
Phage display is a laboratory technique for the study of protein–protein, protein–peptide, and protein–DNA interactions that uses bacteriophages (viruses that infect bacteria) to connect proteins with the genetic information that encodes ...
, ribosome display, bacterial display, and mRNA display
mRNA display is a display technique used for ''in vitro'' protein, and/or peptide evolution to create molecules that can bind to a desired target. The process results in translated peptides or proteins that are associated with their mRNA progenitor ...
.
References
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).
*
{{Protein methods
Display techniques