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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/30584

Title: The Generation of Affinity Reagents Using High-throughput Phage Display and Building the Foundations of a Novel High-throughput Intrabody Pipeline
Authors: Economopoulos, Nicolas
Advisor: Sidhu, Sachdev
Department: Molecular and Medical Genetics
Keywords: phage display
high-throughput
intrabody
antibody
intrabodies
antibodies
Issue Date: 7-Dec-2011
Abstract: Phage display technology has emerged as the dominant approach in antibody engineering. Here I describe my work in developing a high-throughput method of reliably generating intracellular antibodies. In my first data chapter, I present the first known high-throughput pipeline for antibody-phage display libraries of synthetic diversity and I demonstrate how increasing the scale of both target production and library selection still results in the capture of antibodies to over 50% of targets. In my second data chapter, I present the construction and validation of a novel scFv-phage library that will serve as the first step in my proposed intrabody pipeline. Antibodies obtained from this library will be screened for functionality using a novel yeast-two-hybrid approach and have numerous downstream applications. This high-throughput pipeline is amenable to automation and can be scaled up to thousands of domains, resulting in the potential generation of many novel therapeutic reagents.
URI: http://hdl.handle.net/1807/30584
Appears in Collections:Master

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