A microengraving method for rapid selection of single cells producing antigen-specific antibodies

Nat Biotechnol. 2006 Jun;24(6):703-7. doi: 10.1038/nbt1210. Epub 2006 May 14.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies / isolation & purification*
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cells, Cultured
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods*
  • Hybridomas / cytology*
  • Hybridomas / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods*
  • Surface Properties

Substances

  • Antibodies