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Sci Rep. 2019 Jan 29;9(1):842. doi: 10.1038/s41598-018-37462-1.

A rapid in vitro methodology for simultaneous target discovery and antibody generation against functional cell subpopulations.

Author information

1
Donnelly Centre, University of Toronto, Toronto, M5S 3E1, Canada.
2
Department of Molecular Genetics, University of Toronto, Toronto, M5S 1A8, Canada.
3
Stem Cell and Cancer Research Institute, McMaster University, Hamilton, L8S 4K1, Canada.
4
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, L8N 3Z5, Canada.
5
Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, M5S 3G9, Canada.
6
Princess Margaret Cancer Centre, University Health Network, Toronto, M5G 2MC1, Canada.
7
Department of Bioinformatics and Computational Biology, MD Anderson Cancer Center, Houston, Texas, 77030, USA.
8
Department of Surgery, Faculty of Health Sciences, McMaster University, Hamilton, L8N 3Z5, Canada.
9
Department of Surgery, Toronto General Hospital, Toronto, M5G 2C4, Canada.
10
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, Canada.
11
Donnelly Centre, University of Toronto, Toronto, M5S 3E1, Canada. j.moffat@utoronto.ca.
12
Department of Molecular Genetics, University of Toronto, Toronto, M5S 1A8, Canada. j.moffat@utoronto.ca.
13
Canadian Institute for Advanced Research, Toronto, M5G 1Z8, Canada. j.moffat@utoronto.ca.

Abstract

Cell surface antigen discovery is of great interest for biomedical research both for isolation of rare cell populations and therapeutic targeting. We developed a rapid, cost-effective, fully in vitro technology which facilities the simultaneous target discovery and human antibody generation on the surface of virtually any cell population of interest. We apply our technique to human colorectal cancer-initiating cells (CICs) and identify hundreds of unique human antibodies. We characterized the top three antibody candidates targeting these CICs and identify their protein targets as integrin α7 (ITGA7), HLA-A1 and integrin β6 (ITGB6). We demonstrate that these antibodies can be used to isolate self-renewing colorectal CICs, and that the integrin α7 antibody can prospectively identify glioblastoma brain tumor initiating cells as well as human muscle stem cells. We also demonstrate that genetic ablation of integrin β6 impedes colorectal CIC function. The methodology can be readily applied to other cell populations including stem cells, cancer, or immune cells to facilitate the rapid identification of novel targets and simultaneous generation of potent and specific antibodies with therapeutic potential.

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