Nanowell-Based Immunoassays for Measuring Single-Cell Secretion: Characterization of Transport and Surface Binding

• Main Authors: Torres, Alexis J. (Contributor), Hill, Abby (Contributor), Love, John C (Author)
• Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Chemical Engineering (Contributor), Koch Institute for Integrative Cancer Research at MIT (Contributor), Love, J. Christopher (Contributor)
• Format: Article
• Language: English
• Published: American Chemical Society (ACS), 2016-02-19T02:20:54Z.
• Subjects: Article
• Online Access: Get fulltext

Arrays of subnanoliter wells (nanowells) provide a useful system to isolate single cells and analyze their secreted proteins. Two general approaches have emerged: one that uses open arrays and local capture of secreted proteins, and a second (called microengraving) that relies on closed arrays to capture secreted proteins on a solid substrate, which is subsequently removed from the array. However, the design and operating parameters for efficient capture from these two approaches to analyze single-cell secretion have not been extensively considered. Using numerical simulations, we analyzed the operational envelope for both open and closed formats, as a function of the spatial distribution of capture ligands, their affinities for the protein, and the rates of single-cell secretion. Based on these analyses, we present a modified approach to capture secreted proteins in-well for highly active secreting cells. This simple method for in-well detection should facilitate rapid identification of cell lines with high specific productivities.
National Institutes of Health (U.S.)/National Institute of Allergy and Infectious Diseases (U.S.) (5P01AI045757)
National Cancer Institute (U.S.) (Koch Institute Support (Core) Grant P30-CA14051)