Optimized protocol for antibody yeast display



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Yeast display is an established technology used to discover proteins that have increased binding affinities to their targets. There are many steps within the process that affect the final binding affinity between proteins. The Maynard Lab regularly utilizes display technologies to discover antibodies that bind strongly to targets of interest prior to antibody engineering, in vitro testing, and in vivo studies. For the purpose of efficiency and standardization, the goal of this work is to develop a protocol associated with yeast display for discovery of potent antibodies. In this work, the variable domain is expressed on the surface of yeast cells, in which each cell has a unique heavy chain genotype and matching phenotype based on error-prone mutagenesis. After artificially producing millions of yeast cells with different heavy chain regions, the cells are sorted based on their affinity towards a receptor. DNA sequences that encode for successful binders are used to recombinantly clone, express, and characterize antibodies as leading candidates with increased binding affinity to targets. This revised protocol is lab-specific, wherein details about execution of experiments are contextual to the work done in the Maynard Lab. Every bullet-point and step in this report is a lesson learned from significant testing and troubleshooting.


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