Abstract
Here we report a new method for isolating antigen-specific antibody-secreting cells (ASCs) using a microwell array chip, which offers a rapid, efficient and high-throughput (up to 234,000 individual cells) system for the detection and retrieval of cells that secrete antibodies of interest on a single-cell basis. We arrayed a large population of lymphoid cells containing ASCs from human peripheral blood on microwell array chips and detected spots with secreted antibodies. This protocol can be completed in less than 7 h, including 3 h of cell culture. The method presented here not only has high sensitivity and specificity comparable with enzyme-linked immunospot (ELISPOT) but it also overcomes the limitations of ELISPOT in recovering ASCs that can be used to produce antigen-specific human monoclonal antibodies. This method can also be used to detect cells secreting molecules other than antibodies, such as cytokines, and it provides a tool for cell analysis and clinical diagnosis.
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Acknowledgements
We thank S. Hirota for technical assistance and K. Hata for secretarial work. We are very grateful to M. Isobe and N. Kurosawa for their helpful discussions. This research was supported by grants from the Hokuriku Innovation Cluster for Health Science Project of the Ministry of Education, Culture, Sports, Science and Technology, Japan.
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H.K. conceived of the ISAAC method. H.K. and A.M. supervised the project. A.J. designed and performed experiments. T. Obata developed the chip and A.J. tested it. A.J., T. Ozawa and K.T. tested and showed the utility of the protocol.
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H.K. and A.M. are unsalaried directors of SC World Inc.
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Jin, A., Ozawa, T., Tajiri, K. et al. Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array. Nat Protoc 6, 668–676 (2011). https://doi.org/10.1038/nprot.2011.322
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DOI: https://doi.org/10.1038/nprot.2011.322
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