title: A droplet-based heterogeneous immunoassay for screening single cells secreting antigen-specific antibodies.
abstract: High throughput heterogeneous immunoassays that screen antigen-specific antibody secreting cells are essential to accelerate monoclonal antibody discovery for therapeutic applications. Here, we introduce a heterogeneous single cell immunoassay based on alginate microparticles as permeable cell culture chambers. Using a microfluidic device, we encapsulated single antibody secreting cells in 35-40 μm diameter alginate microbeads. We functionalized the alginate to capture the secreted antibodies inside the microparticles, enabling single cell analysis and preventing the cross-talk between the neighboring encapsulated cells. We demonstrated non-covalent functionalization of alginate microparticles by adding three secondary antibodies to the alginate solution to form high molecular weight complexes that become trapped in the porous nanostructure of alginate and capture the secreted antibodies. We screened anti-TNF-alpha antibody-secreting cells from a mixture of antibody-secreting cells.
authors.affiliation: School of Engineering and Applied Sciences, Harvard University, Cambridge, USA. sakbari@seas.harvard.edu.
authors.forename: Samin
authors.initials: S
authors.lastname: Akbari
authors.forename: Tohid
authors.initials: T
authors.lastname: Pirbodaghi
mesh_terms: D000906:Antibodies / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*
chemical_list: D000906:Antibodies; D000941:Antigens