A microengraving method for rapid selection of single cells producing antigen-specific antibodies.
Monoclonal antibodies that recognize specific antigens of interest are used as therapeutic agents and as tools for biomedical research. Discovering a single monoclonal antibody requires retrieval of an individual hybridoma from polyclonal mixtures of cells producing antibodies with a variety of specificities. The time required to isolate hybridomas by a limiting serial-dilution, however, has restricted the diversity and breadth of available antibodies. Here we present a soft lithographic method based on intaglio printing to generate microarrays comprising the secreted products of single cells. These engraved arrays enable a rapid (<12 h) and high-throughput (>100,000 individual cells) system for identification, recovery and clonal expansion of cells producing antigen-specific antibodies. This method can be adapted, in principle, to detect any secreted product in a multiplexed manner.
Love
J Christopher
JC
Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, Massachusetts 02142, USA. love@cbrinstitute.org
Ronan
Jehnna L
JL
Grotenbreg
Gijsbert M
GM
van der Veen
Annemarthe G
AG
Ploegh
Hidde L
HL
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006825:Hybridomas / Q000166:cytology* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D013499:Surface Properties
D000906:Antibodies