Engineered Discoidin Domain from Factor VIII Binds αvβ3 Integrin with Antibody-like Affinity

Author: Kym, Eugene Yongshik (Gene)

Year: 2014

Degree: Dissertation (Ph.D.)

Advisor: Mayo, Stephen L.

Committee Members: Deshaies, Raymond Joseph; Chan, David C.; Tirrell, David A.; Mayo, Stephen L.

Option: Biology

DOI: 10.7907/PTZH-PB29

Abstract

Alternative scaffolds are non-antibody proteins that can be engineered to bind new targets. They have found useful niches in the therapeutic space due to their smaller size and the ease with which they can be engineered to be bispecific. We sought a new scaffold that could be used for therapeutic ends and chose the C2 discoidin domain of factor VIII, which is well studied and of human origin. Using yeast surface display, we engineered the C2 domain to bind to αvβ3 integrin with a 16 nM affinity while retaining its thermal stability and monomeric nature. We obtained a crystal structure of the engineered domain at 2.1 Å resolution. We have christened this discoidin domain alternative scaffold the “discobody.”

Files

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