Developing cell-specific antibodies to endothelial progenitor cells using avian immune phage display technology
Hanley, S. A.
Taylor, W. R.
Griffin, M. D.
Finlay, W. J. J.
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Bowes, T. Hanley, S. A.; Liew, A.; Eglon, M.; Mashayekhi, K.; O'Kennedy, R.; Barry, F.; Taylor, W. R.; O'Brien, T.; Griffin, M. D.; Finlay, W. J. J.; Greiser, U. (2011). Developing cell-specific antibodies to endothelial progenitor cells using avian immune phage display technology. Journal of Biomolecular Screening 16 (7), 744-754
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. (Journal of Biomolecular Screening. 2011; 16: 744-754)