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Lookup NU author(s): Dr Aaron Liew
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This study aims at generating immune chicken phage display libraries and single-chain antibodies (scFvs) specifically directed against cell surface markers of cultured peripheral blood mononuclear cells (PBMCs) that contain endothelial progenitor cells (EPCs). In contrast to previous approaches that use well-defined recombinant antigens attached to plastic surfaces that may alter the structure of the proteins, the authors describe a method that maintains the cell surface markers on live cells while providing the opportunity to rapidly screen entire libraries for antibodies that bind to unknown cell surface markers of progenitor/stem cells. Chickens immunized with live EPCs, consisting of a heterogeneous population of lymphocytes and monocytes, demonstrated a robust immune response. After three rounds of biopanning, the authors purified and characterized three unique scFvs called UG1-3. Codon-optimized recombinant UG1 (gUG-1) shows binding by flow cytometry to circulating CD14-positive cells in peripheral blood consistent with predominant expression of a target protein on monocyte subsets. The authors describe the successful use of immunization of chickens for the generation of scFvs against a heterogenous population of EPCs displaying unknown cell surface markers and demonstrate the strong potential of phage display technology in the development of reagents for the isolation and characterization of stem/progenitor cells.
Author(s): Bowes T, Hanley SA, Liew A, Eglon M, Mashayekhi K, O'Kennedy R, Barry F, Taylor WR, O'Brien T, Griffin MD, Finlay WJ, Greiser U
Publication type: Article
Publication status: Published
Journal: Journal of Biomolecular Screening
Year: 2011
Volume: 16
Issue: 7
Pages: 744-754
Print publication date: 18/05/2011
ISSN (print): 1087-0571
ISSN (electronic): 1552-454X
Publisher: Sage
URL: http://dx.doi.org/10.1177/1087057111407067
DOI: 10.1177/1087057111407067
PubMed id: 21593485
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