title: Developing cell-specific antibodies to endothelial progenitor cells using avian immune phage display technology.
abstract: 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.
authors: Regenerative Medicine Institute, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
authors: Tyrone
authors: T
authors: Bowes
authors: Shirley A
authors: SA
authors: Hanley
authors: Aaron
authors: A
authors: Liew
authors: Marc
authors: M
authors: Eglon
authors: Kaveh
authors: K
authors: Mashayekhi
authors: Richard
authors: R
authors: O'Kennedy
authors: Frank
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authors: Barry
authors: W Robert
authors: WR
authors: Taylor
authors: Timothy
authors: T
authors: O'Brien
authors: Matthew D
authors: MD
authors: Griffin
authors: William J J
authors: WJ
authors: Finlay
authors: Udo
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authors: Greiser