UIC-2014-141 – Combining Kunkel mutagenesis and phage-display to create bivalent affinity reagents

Megaprimer Shuffling for Tandem Affinity Reagents (MegaSTAR) is a UIC technology that converts the output of affinity selection to a bivalent display format. Bivalent reagents have many applications however due to their labor intensive creation, they have been avoided as a viable solution. UIC researchers have invented a method using phage-display and Kunkel mutagenesis to create bivalent affinity reagents. These molecules bind 2 distinct areas (epitopes) on the same target protein. This novel method allows for the rapid and efficient generation of these reagents in a high throughput manner to theoretically any target. Epitope binning consists of examining pairs of binders (in soluble protein form) to determine if they compete for the same epitope. This invention allows one to examine many different combinations of affinity reagents without the need for pair-wise analysis. This MegaSTAR technology relies on synthesizing a “bivalent library” by first generating a pool of megaprimers from selection output, and annealing them randomly to a bivalent vector. This new “bivalent library” is then used for further affinity selection to identify tandem reagents with the highest affinity. This greatly reduces the time and cost of identifying pairs of reagents that bind to different epitopes on the same target protein. Nelson Grihalde grihalde@otm.uic.edu 312-996-4129