title: Isolation of a recombinant antibody specific for a surface marker of the corneal endothelium by phage display.
abstract: Cell surface antigens are important targets for monoclonal antibodies, but they are often difficult to work with due to their association with the cell membrane. Phage display is a versatile technique that can be applied to generate binders against difficult targets. Here we used antibody phage display to isolate a binder for a rare and specialized cell, the human corneal endothelial cell. The human corneal endothelium is a medically important cell layer; defects in this layer account for about half of all corneal transplants. Despite its importance, no specific antigens have been found to mark this cell type. By panning a phage library directly on human corneal endothelial cells, we isolated an antibody that bound to these cells and not the other types of corneal cells. Subsequently, we identified the antibody's putative target to be CD166 by immunoprecipitation and mass spectrometry. This approach can be used to isolate antibodies against other poorly-characterized cell types, such as stem cells or cancer cells, without any prior knowledge of their discriminating markers.
authors.affiliation: Experimental Therapeutics Centre, 31 Biopolis Way, Nanos level 3, Singapore, 138669 Singapore.
authors.forename: Simone
authors.initials: S
authors.lastname: Dorfmueller
authors.affiliation: Experimental Therapeutics Centre, 31 Biopolis Way, Nanos level 3, Singapore, 138669 Singapore.
authors.forename: Hwee Ching
authors.initials: HC
authors.lastname: Tan
authors.affiliation: Experimental Therapeutics Centre, 31 Biopolis Way, Nanos level 3, Singapore, 138669 Singapore.
authors.forename: Zi Xian
authors.initials: ZX
authors.lastname: Ngoh
authors.affiliation: Experimental Therapeutics Centre, 31 Biopolis Way, Nanos level 3, Singapore, 138669 Singapore.
authors.forename: Kai Yee
authors.initials: KY
authors.lastname: Toh
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Gary
authors.initials: G
authors.lastname: Peh
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Heng-Pei
authors.initials: HP
authors.lastname: Ang
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Xin-Yi
authors.initials: XY
authors.lastname: Seah
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros Level 4, Singapore, 138668 Singapore.
authors.forename: Angela
authors.initials: A
authors.lastname: Chin
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros Level 4, Singapore, 138668 Singapore.|Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore, Singapore.
authors.forename: Andre
authors.initials: A
authors.lastname: Choo
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Jodhbir S
authors.initials: JS
authors.lastname: Mehta
authors.affiliation: Institute of Bioengineering and Nanotechnology, Synthetic Biosystems, 31 Biopolis Way, #04-01 Nanos, Singapore, 138669 Singapore.|Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: William
authors.initials: W
authors.lastname: Sun
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000918:Antibody Specificity; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D015816:Cell Adhesion Molecules, Neuronal / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004728:Endothelium, Corneal / Q000166:cytology / Q000378:metabolism*; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D047468:Immunoprecipitation; D009928:Organ Specificity; D019151:Peptide Library; D061251:Primary Cell Culture; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D020410:Activated-Leukocyte Cell Adhesion Molecule
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015415:Biomarkers; D015816:Cell Adhesion Molecules, Neuronal; D005326:Fetal Proteins; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C105613:ALCAM protein, human; D020410:Activated-Leukocyte Cell Adhesion Molecule