原创力文档discovery of t cell antigens by high-throughput screening of synthetic minigene libraries发现了t细胞抗原合成微基因的高通量筛选库.pdfVIP
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discovery of t cell antigens by high-throughput screening of synthetic minigene libraries发现了t细胞抗原合成微基因的高通量筛选库
Discovery of T Cell Antigens by High-Throughput
Screening of Synthetic Minigene Libraries
1 1 1 1¤ 1
Brian D. Hondowicz , Katharine V. Schwedhelm , Arnold Kas , Michael A. Tasch , Crystal Rawlings ,
1 4 1 1 2
Nirasha Ramchurren , Martin McIntosh , Leonard A. D’Amico , Srinath Sanda , Nathan E. Standifer , Jay
3 1
Shendure , Brad Stone *
1Translational Research Program, Benaroya Research Institute, Seattle, Washington, United States of America, 2 Clinical Immunology, Amgen, Seattle, Washington, United
States of America, 3 Department of Genome Sciences, University of Washington, Seattle, Washington, United States of America, 4 Computational Biology, Fred Hutchinson
Cancer Research Center, Seattle, Washington, United States of America
Abstract
The identification of novel T cell antigens is central to basic and translational research in autoimmunity, tumor immunology,
transplant immunology, and vaccine design for infectious disease. However, current methods for T cell antigen discovery
are low throughput, and fail to explore a wide range of potential antigen-receptor interactions. To overcome these
limitations, we developed a method in which programmable microarrays are used to cost-effectively synthesize complex
libraries of thousands of minigenes that collectively encode the content of hundreds of candidate protein targets. Minigene-
derived mRNA are transfected into autologous antigen presenting cells and used to challenge complex populations of
purified peripheral blood CD8+ T cells in
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