Heterofunctional Particles as Single Cell Sensors to Capture Secreted Immunoglobulins and Isolate Antigen‐Specific Antibody Secreting Cells
Autor: | Doan Nguyen, Troy D. Randall, Todd Sulchek, Ignacio Sanz, Ye Lim Lee, Katily Ramirez, John E. Bradley, So-Yun Han, Shuya Kyu, Frances Eun-Hyung Lee |
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Rok vydání: | 2021 |
Předmět: |
Adult
Cell Biomedical Engineering Pharmaceutical Science chemical and pharmacologic phenomena 02 engineering and technology 010402 general chemistry 01 natural sciences Virus Biomaterials Antigen medicine Humans Secretion Antigens Antibody-Producing Cells Hybridomas biology Chemistry Vaccination Antibodies Monoclonal hemic and immune systems 021001 nanoscience & nanotechnology Molecular biology 0104 chemical sciences 3. Good health Secretory protein medicine.anatomical_structure Antibody-Secreting Cells biology.protein Protein G Antibody 0210 nano-technology |
Zdroj: | Advanced Healthcare Materials. 10:2001947 |
ISSN: | 2192-2659 2192-2640 |
Popis: | Isolating cells based on their secreted proteins remain a challenge. The authors demonstrate a capacity for high throughput single-cell protein secretion analysis and isolation based on heterofunctional particles combined with fluorescence activated cell sorting (FACS). The workflow shows that antibody secreting cells (ASCs) specific for the H1 protein from influenza virus can be isolated from B cells. The workflow consists of incubating anti-CD27 particles with the ASCs, capturing locally secreted immunoglobulins with Protein G on the particles, and identifying immunoglobulins specific to H1 via fluorescent labeled antigens followed by FACS to enrich antigen-specific ASCs. Two particles designs, Janus and mixed, are tested with hybridoma cells. Mixed particles are found to improve antibody collection, while Janus particles are found to bind target cells more effectively. Targeted hybridoma cells in coculture with non-specific hybridoma cells are identified with a sensitivity of 96% and specificity of 98%. Heterofunctional particles are used to capture ASCs that secrete antibodies specific for influenza virus from B cells from healthy adults isolated from blood after vaccination. Positive H1-tetramer sorted ASCs are validated using single ASC cultures and identify 23/56 cells specific for H1 demonstrating 164-fold enrichment from total B cells and 14.6-fold enrichment from total ASCs. |
Databáze: | OpenAIRE |
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