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

Nature Protocols volume 6, pages 509–522 (2011)Cite this article

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Abstract

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.

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Figure 1: Flowchart of microselection.
Figure 2: Identification and pictures of target cell position in Step 12.

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Acknowledgements

We thank The Danish Council for Independent Research—Technology and Production Sciences (09-065063) and the FP6 EU project, PROTEOMAGE (LSHM-CT-2005-518230), for financial support. FCMB is thanked for partial financing of M.D.S.

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Authors and Affiliations

  1. Department of Molecular Biology, Aarhus University, Aarhus, Denmark

    Morten Dræby Sørensen & Peter Kristensen

  2. Aarhus School of Engineering, Aarhus University, Aarhus, Denmark

    Peter Kristensen

Authors
  1. Morten Dræby Sørensen
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  2. Peter Kristensen
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Contributions

M.D.S. designed and conducted the experiments in the development of the method and wrote the manuscript. P.K. designed experiments, supervised the project and contributed to the writing of the manuscript. Both authors discussed the results and implications and commented on the manuscript at all stages.

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Correspondence to Peter Kristensen.

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The authors declare no competing financial interests.

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Sørensen, M., Kristensen, P. Selection of antibodies against a single rare cell present in a heterogeneous population using phage display. Nat Protoc 6, 509–522 (2011). https://doi.org/10.1038/nprot.2011.311

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