Yeast Surface Display Library Screening Service
Alpha Lifetech specializes in the yeast display library construction and screening of antibody libraries, offering the capability to generate different forms of yeast libraries and screen them for high affinity and specific antibodies for clients worldwide. With a team of expert researchers, advanced technology and equipment, we can construct yeast libraries targeting a wide range of disease-related proteins. We can provide phage display and yeast display technologies, including protein yeast display, small molecule yeast display, antibody yeast display, etc
Our yeast display libraries have large capacity and high diversity, which provide a high foundation for the effective screening of specific antibody sequences. We can construct different forms of antibody yeast display libraries (IgG, scFv, VHH, Fab antibody libraries), protein libraries, peptide libraries, cDNA libraries, etc. according to your needs. Additionally, we can also develop antibody libraries of various properties, including immune libraries, natural libraries, synthetic libraries, semi-synthetic libraries, and disease antibody libraries.
Introduction to Yeast Display System
Yeast surface display technology is a method of displaying recombinant proteins on the surface of yeast through gene fusion. The most common yeast display system uses the target protein fused to the C-terminus of the Aga2p subunit of the alpha lectin mating protein, mainly consisting of epitope tags on both sides of the target protein: the N-terminal 9-amino acid hemagglutinin (HA) tag and the C-terminus 10 amino acid c-myc tag. The 69 amino acid Aga2p subunit binds to the 725 amino acid alpha lectin Aga1p subunit via two disulfide bonds, and Aga1p is anchored to the cell wall via a β 1,6-glucan covalent bond. Therefore, the target protein is displayed on the surface of yeast cells and subsequently recognized by the corresponding ligand. Separate functional proteins from libraries through flow cytometry screening.

Fig 1: Principle of yeast surface display. (Figure source: Applications of Yeast Surface Display for Protein Engineering.)
Introduction to Yeast Surface Display Library
Yeast display sorting is based on yeast surface display, mainly used to screen antibody libraries targeting cell surface proteins. Due to the low non-specific binding between yeast cells and other cell surfaces, yeast biological selection can screen large binding libraries to find rare clones.
Yeast Surface Display Library Screening Service
Prepare plasmids and culture yeast cells, synthesize DNA encoding the target protein and clone it into a yeast expression vector containing inducible promoters, signal peptides, and target genes fused to surface display proteins (such as Aga2p). The gene encoding the target protein can be fused with the gene encoding the yeast cell wall protein (usually Aga2p), and the fused gene can be transformed into yeast cells through electroporation. After transformation, yeast cells expressing antibody genes on their surface can undergo antigen-specific screening tests: the yeast library is incubated with the target antigen and yeast cells that specifically bind to it are selected. Using fluorescence activated cell sorting (FACS) to isolate yeast cells displaying proteins with desired characteristics, yeast display library screening can be performed with a maximum library size of~10 ^ 8-10 ^ 9 yeast cells. Positive clones can then be isolated for further analysis or downstream applications. Once specific antibody clones are identified from the antibody library, they can be further characterized and mass-produced, and purified using techniques such as protein A/G affinity chromatography to complete the entire process of yeast display screening and antibody production.
Process of Yeast Display Library Screening

Fig.2 Yeast display library screening process
Yeast Display Library Screening Service Workflow
Steps | Service Content | Timeline |
---|---|---|
Antigen Preparation | Antigen type: If the customer can provide antigens, corresponding samples need to be delivered according to the type: recombinant proteins require 3-3.5mg with a purity requirement of over 85%, small molecules need to be conjugated with a purity requirement of over 90%, peptide synthesis needs to be conjugated with a purity requirement of over 90%, sample types such as viruses need to be inactivated, RNA need to be inspected to prevent degradation, and the above antigen types can also be customized for synthesis. | 2-3 weeks |
Animal Immunity | The number of animal immunizations is 5, and it is necessary to determine whether to increase the number of immunizations based on serum titer testing. Antigen immunity: protein/virus antigen potency>105; Peptide/small molecule antigen potency>10^4 | 5-6 weeks |
Template cDNA Preparation | Separate plasma PBMCs, extract total RNA (RNA extraction kit) and reverse to cDNA. | 1 day |
Library Construction | Using library cDNA as a template, VHH gene was amplified by two rounds of PCR, and VHH gene splicing yeast display vector was constructed. The vector was transformed into yeast cells by electroporation to construct an antibody library. Randomly select 48 clones and identify the positive rate (>90%) using PCR method; Calculate library capacity (10^7-10^8), NGS sequencing to determine correct library insertion rate (>90%) and library diversity. | 2 weeks |
Library Screening | Default three rounds of screening: Fluorescence labeled protein FACS screening, followed by NGS sequencing in the third round. Positive clones were selected for single gram induction expression and ELISA detection. All positive clones were selected for gene sequencing, and different CDR region sequences were selected. | 2-3 weeks |
Antibody Verification | Constructing appropriate expression vectors for antibody sequences can facilitate antibody expression, antibody purification, ELISA and BLI validation of antibody antigen binding to verify antibody affinity, and flow cytometry blockade to validate cell function. | 1 week |
Case of Yeast Surface Display Library Screening
In the literature of yeast surface display platform for rapid discovery of conformationally selective nanobodies, the authors established a complete in vitro nanobody discovery platform based on yeast surface display. Firstly, a synthetic nanobody library was designed starting from camel genes. Figure D shows that the nanobody has an HA tag at the carboxyl end, and then the nanobody is covalently fixed on the yeast cell wall. Figure E shows the nanobody screening process. Yeast with antigen affinity nanobodies were isolated, amplified, and repeatedly selected for antibodies by FACS. The authors discovered conformational selective nanobodies targeting two different human GPCRs through this platform.

Fig 3: Design and construction of synthetic nanobody library. (Figure source: Yeast surface display platform for rapid discovery of conformationally selective nanobodies.)
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