A droplet-based heterogeneous immunoassay for screening single cells secreting antigen-specific antibodies

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 mu m diameter alginate microbeads. We functionalized the alginate to capture the secreted antibodies inside the microparticles, enabling single cell analysis and preventing the crosstalk 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.


Publié dans:
Lab On A Chip, 14, 17, 3275-3280
Année
2014
Publisher:
Cambridge, Royal Soc Chemistry
ISSN:
1473-0197
Laboratoires:




 Notice créée le 2014-10-23, modifiée le 2018-12-03


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