Vectors for Yeast Display

PYDL Yeast Display Vector Series

Next-Generation Yeast Display Vectors for Antibody and Protein Engineering.

The PYDL Vector Series is a line of plasmids dedicated to Yeast Display, a technology renowned for revolutionizing protein engineering and antibody discovery. Leveraging our expertise in molecular biology, these vectors provide a robust and versatile platform for displaying diverse protein libraries on the surface of Saccharomyces cerevisiae cells, enabling rapid screening and selection of variants with desired properties.

Overview of Yeast Display Technology

Yeast display is a powerful method where engineered proteins are presented on the surface of S. cerevisiae cells, allowing for high-throughput screening.

Key Advantages

  • Eukaryotic Expression: Proteins are folded and modified in a native-like eukaryotic environment, increasing the chance of functional display for complex proteins like antibodies or receptors.

  • Quantitative Analysis: Flow cytometry enables highly sensitive, quantitative, and high-throughput screening of library members, facilitating the isolation of rare high-affinity binders.

  • Facile Genetic Manipulation: Yeast is genetically tractable, which simplifies library construction and mutagenesis.

  • Direct Linkage: The displayed protein is genetically linked to its encoding DNA, simplifying the recovery and sequencing of selected clone.

  • Versatility: The system is suitable for displaying a wide range of proteins, including scFvs and VHHs (single-domain antibodies).

How it Works

The system utilizes the native mating agglutination machinery, specifically the Aga1p/Aga2p components of S. cerevisiae.

  1. The protein of interest (POI) is expressed as a fusion protein with Aga2p in yeast strains (like EBY100) that also express Aga1p.

  2. The Aga2p-POI fusion forms two critical disulfide bonds with the larger, cell-wall-anchored component, Aga1p.

  3. This covalent linkage creates a stable, display complex that anchors the POI to the yeast cell surface, making it accessible for directed evolution, binding assays, and flow cytometry screening.

Figure 1. Yeast surface display. Representation of N- and C-terminal Aga2p fusions for display of an scFv (as an example) on yeast. Surface display is detected by antibody staining of epitope tag (HA, or similar) and target binding using biotinylated antigen followed by fluorescently-labeled streptavidin.

pYDL™ Vectors

The pYDL vectors facilitate yeast display by providing tools for display in N-terminal and C-terminal Aga2p fusion orientations with a display epitope tag of choice, and a tryptophan selection marker for isolating transformed auxotrophic yeast. These vectors utilize a CEN/ARS origin of replication to maintain a single stable copy per cell without chromosomal integration thus maintaining the genotype-phenotype linkage required for quality yeast display libraries.

Vector

Catalog Number

Display Orientation

Epitope Tag

pYDL-12

YD0100

N-terminal

HA

pYDL-13

YD0101

N-terminal

Myc

pYDL-22

YD0102

C-terminal

HA

pYDL-23

YD0103

C-terminal

Myc

Table 1. List of pYDL series vectors

Key Features of pYDL Vectors

  • Display Orientation: Offers both N-terminal and C-terminal Aga2p fusion orientations to minimize steric hindrance and optimize the functional display of your protein of interest (POI).

  • Epitope Tags: Includes a display epitope tag (HA or c-Myc) of your choice for surface display detection.

  • Yeast Selection: Contains a tryptophan selection marker for isolating transformed auxotrophic yeast.

  • Replication: Utilizes a CEN/ARS origin of replication to maintain a single stable copy per cell without chromosomal integration, which is essential for preserving the genotype-phenotype linkage required for high-quality yeast display libraries.

  • Promoter: Expression is controlled by the GAL1 promoter.

  • Performance Data: Flow cytometry analysis demonstrates that all four pYDL vectors successfully enable both single-domain antibody surface display and antigen binding.

Figure 2. pYDL series vector comparison. Flow cytometry analysis of display and binding of a single-domain antibody to its antigen using various pYDL vectors.

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