Biological Probes

Antibodies are among the most commonly used research tools in biomedicine. They represent key tools to detect presence of proteins; to monitor protein localization in cells and tissues; and to study protein-protein interaction and signaling networks. Despite their importance, and the availability of the entire human genome sequence for nearly ten years, there does not exist a library of high-quality antibodies to each human protein. Access to research tools, such as antibodies, for novel proteins is a limiting factor in our exploration of the human genome and in our understanding of disease. In fact, most biological research is still centered on genes and proteins that were known already at the time of the completion of the human genome project. In part this is due to the lack of appropriate research tools. Consequently generation of high-quality, ideally renewable, antibodies to novel protein will have a great impact to enable and stimulate research on novel proteins.

Project objectives and scope

The direct goal of this project is to develop 200 high-quality renewable antibodies to proteins implicated in epigenetic events. On a larger scale this aims to provide the scientific community with research tools that enable in-depth research to better understand the roles and functions of the targeted proteins in health and disease. The project is carried out with additional financial support from Life Technologies. 

Antibody generation - background

Many projects have attempted large scale generation of antibodies. These have revealed three major challenges associated with creating renewable antibodies for large numbers of human proteins: 1) the availability of high-quality antigens; 2) the need to manage the process effectively: and 3) the lack of generally-accepted quality-control criteria (Blow Nature 2007; Schofield et al. Genome Biol 2007).

The SGC has the ability to produce large numbers of proteins of high quality and has the experience in managing large-scale projects. During the course of a pilot project  the SGC worked with the community to identify technologies and partners for a larger scale project. The pilot project demonstrated that in-vitro phage display methods are very well suited for systematic antibody generation using SGC proteins as antigens

Research Plan

The project is carried out as a close collaboration with three of the world leaders in the field of phage display selection technologies (Tony Kossiakoff and Shohei Koide at University of Chicago, and Sachdev Sidhu at the University of Toronto), the laboratory of Jack Greenblatt at University of Toronto, and with Life Technologies.

Antigens are produced using the SGC protein production platform at the University of Toronto under the leadership of Dr. Susanne Gräslund and provided to one or more of the recombinant affinity reagent producers. Candidate antibodies are validated to meet quality criteria adapted to different applications. This process has been accomplished in its entirety as part of the mentioned pilot project (Colwill et al. Nature Methods 2011).