Developing Cell-Specific Antibodies to Endothelial Progenitor Cells Using Avian Immune Phage Display Technology

作者：Bowes Tyrone; Hanley Shirley A; Liew Aaron; Eglon Marc; Mashayekhi Kaveh; O'Kennedy Richard; Barry Frank; Taylor W Robert; O'Brien Timothy; Griffin Matthew D; Finlay William J J; Greiser Udo^*

来源：Journal of Biomolecular Screening, 2011, 16(7): 744-754.

DOI：10.1177/1087057111407067

• phage display
• single-chain antibodies
• endothelial progenitor cells
• cell surface markers
• vasculature

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)

• 出版日期2011-8