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Microselection – affinity selecting antibodies against a single rare cell in a heterogeneous population
Authors:Morten Dræby Sørensen  Inge Errebo Agerholm  Britta Christensen  Steen Kølvraa  Peter Kristensen
Institution:aDepartment of Molecular Biology, University of Aarhus, Aarhus, Denmark;bFCMB Aps, Vejle, Denmark;cThe Fertility Clinic, Brædstrup Hospital, Denmark;dDepartment of Clinical Genetics, Vejle Hospital, IRS, University of Southern Denmark, Odense, Denmark
Abstract:Rare cells not normally present in the peripheral bloodstream, such as circulating tumour cells, have potential applications for development of non‐invasive methods for diagnostics or follow up. Obtaining these cells however require some means of discrimination, achievable by cell type specific antibodies. Here we have generated a microselection method allowing antibody selection, by phage display, targeting a single cell in a heterogeneous population. One K562 cell (female origin) was positioned on glass slide among millions of lymphocytes from male donor, identifying the K562 cell by FISH (XX). Several single cell selections were performed on such individual slides. The phage particles bound to the target cell is protected by a minute disc, while inactivating all remaining phage by UV‐irradiation; leaving only the phage bound to the target cell viable. We hereby retrieved up to eight antibodies per single cell selection, including three highly K562 cell type specific.
Keywords:microselection  single cell selection  phage display  antibody selection  K562  UV irradiation  shadow‐stick  rare cell detection  FISH
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