Protein expression of ARH3-binding partners and purification of two antibodies (Macro-3-FC and Macro(AF1521)-FC)

Abstract

By Linh Tran, Biochemistry

Advisor: In-kwon Kim

Presentation ID: PM_D02

Abstract: The first part of this project is protein test expression, which is proceeded to produce specific proteins on E.coli cell lines. Production of these recombinant proteins is important for genetic engineering, pharmacology and other scientific fields. The procedure includes transformation, cloning, inducing with IPTG and testing protein expression by running electrophoresis gels. The proteins need to be soluble to fully function in cell. Therefore, a positive result of gel electrophoresis is when the specific protein shows up in the IS lane (induced and soluble.) These proteins are considered as binding partners of ARH3, which is a serine mono-ADP-ribosylhydrolase. ARH3 contributes to the regulation of the basal and stress-induced ADP-ribosylome enzyme, which involve in post-translational modification in cells. Finding protein partners for ARH3 contributes to the study of how ARH3 protein is functionally regulated. Some further studies of ARH3 protein partners would be the mechanism of how these proteins work, how they are regulated and how they interact with ARH3 protein. The second part of the project is purification of two antibodies: Macro-3-FC and Macro(AF1521)-FC. They are monoclonal antibodies which have some significant application in pharmaceutical industry. The purification methods include Nickel column, ion exchange and size exclusion chromatography.