Developing cell-specific antibodies to endothelial progenitor cells using avian immune phage display technology

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.
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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)
SAGE Publications
Publisher DOI
Attribution-NonCommercial-NoDerivs 3.0 Ireland