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Platform

Yeast Surface Display

Novel Yeast display-based single domain antibody discovery platform 

Yeast display or yeast surface display is a novel technique widely used to express the proteins at the yeast surface after translation and maturation in a eukaryotic system. Thus, the yeast display is superior to phage display for display of protein or antibody because the eukaryotic expression environment of yeast cells ensures fine tuning such as folding, modification, and translocation prior to display on the surface. NBbiolab has developed yeast display technology for nanobody discovery , affinity maturation, etc.


Yeast surface display (or simply, yeast display) is a potent protein engineering tool for a broad range of biotechnology and biomedical applications and is widely used for in vitro protein evolution. Yeast surface display has been regarded as an alternative method for isolating and engineering antibody fragments (Fab, scFv) from immune and non-immune libraries, and has been applied to isolate recombinant antibodies with binding specificity to a variety of proteins, peptides, small molecules and T cell receptors.

In a general “a-agglutinin” display system, the protein of interest  be fused to the C-terminus of the a-agglutinin Aga2p subunit. Following translation, the 69-amino acid Aga2p subunit associates with 725-amino acid a-agglutinin Aga1p subunit via two disulfide bonds. The fusion protein is subsequently secreted to the extracellular space where Aga1p is anchored to the cell wall via a β1,6-glucan covalent linkage. As a result, the protein of interest is displayed on the cell surface where it is accessible by soluble ligands (as shown below).

Yeast is superior to phage for nanobody display in that the eukaryotic expression environment of yeast cells is more suitable for  protein folding, modification, and translocation for display. Therefore, the yeast displayed antibodies are conformationally and functionally closer to  native counterparts.


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Fig.1.  Schematic representation of yeast surface display 


Benifits

  1. Yeast has a secretory pathway similar to that of eukaryotes. Proteins are folded in endoplasmic reticulum, in which chaperones, folding enzymes and quality control mechanisms ensure that only correctly folded proteins are secreted. Phage is prokaryotic system, some antibodies clones are toxic to E. coli , its growth is slow or even die, resulting in Library bias and clones loss.

  2.  When screening antibodies with higher affinity through phage display, it is usually adversely affected by the screening process, not only by affinity, but also by the level of antibody expression. The yeast display system adopts FACS sorting technology, which is based on antibody affinity and display level, so it can eliminate the deviation caused by expression and distinguish clones with small affinity.

  3. Yeast display is a multivalent display, which can directly select high, medium low affinity clones through sorting at a time. It is very convenient.

  4.  By using FACS with double staining, antibody affinity can be directly determined on the surface of yeast cells, so it does not need time-consuming subcloning, expression and purification.

  5. The antibody screening speed is fast, and 100000 cells can be sorted in 3 minutes, and 100 + candidate clones can be obtained.

  6.  In the same display system, the affinity and stability of antibody can be optimized.

Yeast Display Service

    With extensive experience , NBbiolab yeast display technology is a highly robust display system for the discovery and optimization of nanobody, , our scientists can assist you in      the most comprehensive services and products based on yeast display technology, which including:

    Yeast Display Library Construction Service

    Yeast Display Library Screening Service

    Yeast Display-Based Antibody Affinity Maturation Service

    If you are interested in learning more about yeast display service, please contact us for more details.

References

1.McMahon, C., Baier, A.S., Pascolutti, R.et al.Yeast surface display platform for rapid discovery of conformationally selective nanobodies.Nat Struct Mol Biol 25, 289–296 (2018) doi:10.1038/s41594-018-0028-6

2.Uchański, T., Zögg, T., Yin, J.et al.An improved yeast surface display platform for the screening of nanobody immune libraries.Sci Rep9,382 (2019) doi:10.1038/s41598-018-37212-3

3.Zhuo Yang,Yue Wan.et al.A cell–cell interaction format for selection of high-affinity antibodies to membrane proteins.PNAS July 23, 2019 116 (30) 14971-14978. doi.org/ 10.1073/pnas.1908571116


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