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Gold Nanoparticle-Based In Vitro Screening on Immunogenic Peptides With T Cells
Yang Yanlin1, Shi Chuanyin2,3, Qian Qijun2, 3, 4*
1Xin Yuan Institute of Medicine and Biotechnology, College of Life Science, Zhejiang Science and Technoloy University, Hangzhou 310018, China;
2Shanghai Cell Therapy Research Institute, Shanghai 201805, China;
3Shanghai Engineering Research Center of Cell Therapy, Shanghai 201805, China;
4Department of Biotherapy, Eastern Hepatobiliary Surgery Hospital, Shanghai 201805, China
2Shanghai Cell Therapy Research Institute, Shanghai 201805, China;
3Shanghai Engineering Research Center of Cell Therapy, Shanghai 201805, China;
4Department of Biotherapy, Eastern Hepatobiliary Surgery Hospital, Shanghai 201805, China
Abstract: The core of highly-anticipated immunotherapy lies on the epitope which can lead to activated immune response. This study employed gold nanoparticles (AuNP) as panning carrier to conjugate tumor antigens and incubated with peripheral blood mononuclear cell. Candidates for immunogenic peptides were screened by enzyme-linked immune spots (ELISPOT) assay. Spectrum analysis showed that synthesized peptides can be successfully conjugated to gold nanoparticles. Fluorescence microscopy and transmission electron microscopy demonstrated cellular uptake of conjugation complex. Through the IFN-γ secretion analysis from activated T cells， immunogenicity of different peptides can thus be evaluated. This study optimized experiment parameters such as pH， buffer system， and incubation time etc; shortened the total experiment time from peptide synthesis to immunogenicity analysis; down-scaled to reaction system into 96-well plate and achieved the goal of simple and rapid screening of immunogenic peptide.
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