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Yeast Display-Based Antibody Affinity Maturation Service

Yeast surface display is an effective tool for antibody affinity maturation because yeast can be used as an all-in-one workhorse to assemble, display and screen diversified antibody libraries. Creative Biolabs has developed a pipeline for antibody in vitro affinity maturation based on yeast display system with the principles of mutation and selection. Our affinity yeast display-based maturation service has successfully been used to optimize antibodies for our worldwide clients.

Yeast Display-Based Antibody Affinity Maturation

The affinity of antibodies for their target binding partner is a vital parameter that regulates the biological function of the bound complex. High binding affinity is a desired characteristic of antibodies used for research, therapeutic, and diagnostic applications. Therefore, multiple strategies for increasing protein affinity (termed “affinity maturation”) have been developed, with the most common involving directed evolution and molecular display technologies. In the past few years, yeast display has become a leading platform for affinity maturation; in addition to the above-mentioned advantages, yeast display can distinguish between proteins with only 2-fold differences in affinity, further indicating the sensitivity of this method.

Strategies

A general strategy of affinity maturation via yeast display involves library creation on the order of protein variants by random mutagenesis, followed by the display of these variants on the surface of yeast as fusions to the Aga2p cell wall protein. Subsequently, two main strategies are used to label the yeast-displayed library prior to sorting by FACS. For the first approach, library members are screened based on their equilibrium dissociation constants (KD). And the second approach uses kinetic competition and screens yeast-displayed variants based on their dissociation rate constants. No matter which method is used, high-affinity variants are selected using FACS to isolate cells that exhibit high levels of binding for a given amount of cell surface expression, as measured by a fluorescent antibody against the C-terminal epitope tag and a fluorescently labeled soluble ligand, respectively.

Yeast Display-Based Antibody Affinity Maturation ServiceFig 1. Isolating high-affinity protein variants from a yeast-displayed library by FACS (Cherf and Cochran, 2015).

What We Offer

Yeast antibody display libraries can be coupled with FACS to select high-affinity antibodies that bind specifically to the targeted antigen via labeled target antigens. Application of yeast antibody display combined with FACS allows a more controlled and quantitative selection process that can be fine-tuned to enrich clones with particular binding properties more effectively than phage display. As a world leader in the industry of antibody discovery, Creative Biolabs has extensive experience in yeast display-based antibody affinity maturation. What’s more, with decades of experience in bioassay, we are able to routinely isolate antibodies with equivalent binding specificities to the original antibody and with picomolar binding affinities.

If you are interested in learning more about Creative Biolabs’ yeast display platform, please contact us for more details.

Reference

  1. Cherf, G. M.; Cochran, J. R. Applications of yeast surface display for protein engineering. Yeast Surface Display: Methods, Protocols, and Applications. 2015, 155-175.

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