Selection of antibodies against a single rare cell present in a heterogeneous population using phage display

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  • Morten Dræby Sørensen, Danmark
  • Peter Kristensen, Danmark
Here we describe a new method applying phage-displayed antibody libraries to the selection of antibodies against a single identified cell on a glass slide. This is the only described method that has successfully achieved selection of antibodies against a single rare cell in a heterogeneous population of cells. The phage library is incubated with the slide containing the identified rare cell of interest; incubation is followed by UV irradiation while protecting the target cell with a minute disc. The UV light inactivates all phages outside the shielded area by cross-linking the DNA constituting their genomes. The expected yield is between one and ten phage particles from a single cell selection. The encoded antibodies are subsequently produced monoclonally and tested for specificity. This method can be applied within a week to carry out ten or more individual cell selections. Including subsequent testing of antibody specificity, a specific antibody can be identified within 2 months.
OriginalsprogEngelsk
TidsskriftNature Protocols (Print)
Vol/bind6
Nummer4
Sider (fra-til)509-522
Antal sider14
ISSN1754-2189
DOI
StatusUdgivet - 1 apr. 2011

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