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

Nixon, Allison M L; Duque, Alejandro; Yelle, Nicholas; McLaughlin, Megan; Davoudi, Sadegh; Pedley, Nicolas M; Haynes, Jennifer; Brown, Kevin R; Pan, James; Hart, Traver; Gilbert, Penney M; Singh, Sheila K; O'Brien, Catherine A; Sidhu, Sachdev S; Moffat, Jason.
Sci Rep; 9(1): 842, 2019 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-30696911
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 ß6 (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 ß6 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.
Selo DaSilva