Yeast Display Library Screening Introduction

Yeast surface display is a powerful technology. It has a wide range of applications in various fields, such as protein library screening, protein directed evolution, high affinity antibody sorting, antigen/antibody library construction, affinity maturation, and vaccine production. Yeast display library construction and screening are key steps in yeast display technology. Based on our rich field experience and yeast display platform, Creative Biolabs provides comprehensive services to support yeast display library screening.

Yeast Display Library Screening

Yeast display technology has an important role in antibody development. Yeast display technology is effective for human monoclonal antibody identification and screening of high affinity or high stability antibody. For example, in antibody engineering, the characteristics of yeast eukaryotic expression system are utilized to express antibody library proteins on the surface of yeast cells, which are screened using magnetic beads and flow cytometric sorting technology to obtain high affinity or high stability antibodies. In addition, yeast display-based T cell receptor engineering plays an important role in cell therapy development.

Although a variety of yeast cell lines and a variety of cell wall anchors have been realized for the display of various proteins, the most commonly used is Saccharomyces cerevisiae α-agglutinin. α-agglutinin consists of 2 core subunits, Aga1 and Aga2. In the Saccharomyces cerevisiae α-agglutinin system, the gene encoding the Aga1 protein is stably integrated into the yeast chromosome. However, the gene encoding the protein scaffold-AgA2 fusion protein was cloned into the circular yeast display plasmid vector and was selectively maintained in yeast by nutritional markers. Upon induction of protein expression, the protein scaffold is expressed as a fusion with the Aga2 protein, and the fusion Aga2 protein is then covalently linked to the α-agglutinin Aga1 protein via two disulfide bonds to form a covalent complex that is secreted to the surface of the yeast cell. Expression of both Aga1 and Aga2 is controlled by a galactose-inducible promoter. Because surface display is conditional rather than constitutive, potentially cytotoxic protein scaffolds can be expressed, thus avoiding library bias due to negative selection for such scaffolds. Approximately 50,000 protein-Aga2 fusions can be displayed per yeast cell, and the level of display may be low or high depending on the stability and solubility of the protein scaffold.

Yeast display library screening is a key step in yeast display technology. In the yeast display system, gene expression is regulated by promoters, and secretion efficiency is affected by secretion signals. In addition, the accessibility and activity of the displayed proteins are affected by the length of the anchoring proteins. The ideal combination of promoter, secretion signal, and anchoring protein depends on the target protein. Therefore, the appropriate screening steps are crucial for the results of yeast display technology. Scientists are also continuously optimizing the screening of yeast display libraries for better screening results.

For yeast library screening, the usual experience is to use flow cytometric sorting or magnetic bead screening combined with flow sorting. The immune libraries usually require only 1 round of magnetic bead screening combined with 1-2 rounds of flow cytometric sorting. Multiple antibody sequences with rich diversity and high affinity can be obtained by NGS sequencing, and multiple clones can be further characterized. Multiple rounds of affinity maturation can then be performed as required to improve performance.

Creative Biolabs has a wealth of knowledge and experience in yeast display. We would be happy to share with you our knowledge and experience on yeast display library screening.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

	USA: Europe: Germany:
	Call us at: USA: UK: Germany:
	Fax:
	Email: