Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array
Aishun Jin1, Tatsuhiko Ozawa, Kazuto Tajiri
1Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
Insights
Researchers developed a novel microwell array chip for rapid, high-throughput isolation of antigen-specific antibody-secreting cells (ASCs). This method enables single-cell detection and retrieval, outperforming traditional techniques for antibody discovery and cell analysis.
Area of Science:
- Immunology
- Biotechnology
- Cell Biology
Background:
- Isolating antigen-specific antibody-secreting cells (ASCs) is crucial for therapeutic antibody development.
- Existing methods like ELISPOT have limitations in cell recovery and retrieval for further analysis.
- A need exists for rapid, efficient, and high-throughput methods for single-cell ASC detection and recovery.
Purpose of the Study:
- To develop and validate a novel microwell array chip system for isolating antigen-specific ASCs.
- To enable high-throughput, single-cell detection and retrieval of antibody-secreting cells.
- To provide a method with sensitivity and specificity comparable to ELISPOT but with improved cell recovery.
Main Methods:
- Arraying large populations of lymphoid cells from human peripheral blood onto microwell array chips.
- Detecting spots corresponding to secreted antibodies on a single-cell basis.
- Utilizing a protocol completed in under 7 hours, including cell culture.
Main Results:
- The microwell array chip system allows for rapid and efficient detection and retrieval of antigen-specific ASCs.
- The system supports high-throughput analysis, accommodating up to 234,000 individual cells.
- Sensitivity and specificity are comparable to enzyme-linked immunospot (ELISPOT), with enhanced ASC recovery.
Conclusions:
- The developed microwell array chip method offers a significant advancement for isolating antigen-specific ASCs.
- This technology facilitates the production of antigen-specific human monoclonal antibodies.
- The method is versatile, applicable to detecting other secreted molecules like cytokines, and useful for cell analysis and clinical diagnosis.
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.
