UltraffTM Yeast Display Technology Used in Antibody Discovery and Affinity Maturation

Protein engineering has now become a huge promising industry that attracts the attention of the whole biological world. There are many branches in it. Yeast display is the one that has to be mentioned.

It is a technique that has been wildly used in the field of protein engineering. Professor K. Dane Wittrup and his team first published this technique in their laboratory. Later on, it was sold to Abbott Laboratories in the year of 2001. From the perspective of time, it is still a comparatively new one in the field of protein engineering.

A protein of interest is displayed as a fusion to the Aga2p protein on the surface of yeast. This special type of protein is naturally used by yeast to mediate the contacts between cells during the mating of a yeast cell, by which display of a protein via Aga2p projects the protein away from the cell surface. This procedure has minimized the potential interaction with other molecules on the yeast cell wall. The use of magnetic separation and flow cytometry in conjunction with a yeast based library is a highly effective method to isolate high affinity protein ligands against nearly any receptor through directed evolution.

This unique sort of protein engineering does much better in the expression and processing of eucaryote, the quality control mechanisms of eukaryotic secretory pathway, minimal avidity effects and quantitative library screening through fluorescent-activated cell sorting (FACS) compared with other in vitro evolution methods. Yeast is a eukaryotic organism that allows for complex post-translational modifications to proteins that no other display libraries are able to provide.

Scientists in Creative Biolabs constructed a fully-human, nonimmune library for de novo discovery of therapeutically relevant antibodies. This library consists of 109 human antibody scFv fragments from 16 donors. All these fragments were cloned and expressed on the yeast surface. They obtained nanomolar-affinity scFvs with the flow cytometry or magnetic bead based methods and the UltraffTM Yeast Display Technology. Without measurable clonal diversity loss, the yeast library can be expanded to 1010 fold, which refers to the indefinite expansion of the libraries. All scFv clones are directly assessed on the yeast cell surface by flow cytometry and immunofluorescent labeling, avoiding the separating steps of expression, subcloning and purification. These have demonstratedthat they can be applied for high-throughput antibody isolation for proteomic study.

In addition, Creative Biolabs has created yeast display scFv library HuscL-YDP of the next-generation and fully-human for de novo discovery of therapeutically relevant antibodies. scFv is the most suitable protein to be displayed on yeast surface due to its relatively small size and broad applications. Its library is based on two liters of blood from approximately 20 donors of varying ages and genetic backgrounds. This library consists of over 109 clones can be screened with flow cytometric or magnetic bead based methods in combination with the unique phage technology.

Creative Biolabs is a leading custom service provider that has extensive experience in yeast display. UltraffTM Yeast Display Technology it developed is in widely use of antibody discovery and affinity maturation. Learn more about Creative Biolabs.

Creative Biolabs is a recognized service provider in converting small gene-engineered scFv or Fab human or mouse antibodies derived from phage display antibody library screening.