Please use this identifier to cite or link to this item: https://doi.org/10.1038/srep21661
Title: Isolation of a recombinant antibody specific for a surface marker of the corneal endothelium by phage display
Authors: Dorfmueller, S
Tan, H.C
Ngoh, Z.X
Toh, K.Y
Peh, G 
Ang, H.-P
Seah, X.-Y
Chin, A
Choo, A 
Mehta, J.S 
Sun, W 
Keywords: ALCAM protein, human
biological marker
fetoprotein
leukocyte antigen
monoclonal antibody
nerve cell adhesion molecule
peptide library
protein binding
recombinant protein
single chain fragment variable antibody
antibody specificity
cornea endothelium
cytology
endothelium cell
fibroblast
gene expression
genetics
human
immunoprecipitation
isolation and purification
metabolism
peptide library
primary cell culture
Antibodies, Monoclonal
Antibody Specificity
Antigens, CD
Biomarkers
Cell Adhesion Molecules, Neuronal
Endothelial Cells
Endothelium, Corneal
Fetal Proteins
Fibroblasts
Gene Expression
Humans
Immunoprecipitation
Organ Specificity
Peptide Library
Primary Cell Culture
Protein Binding
Recombinant Proteins
Single-Chain Antibodies
Issue Date: 2016
Citation: Dorfmueller, S, Tan, H.C, Ngoh, Z.X, Toh, K.Y, Peh, G, Ang, H.-P, Seah, X.-Y, Chin, A, Choo, A, Mehta, J.S, Sun, W (2016). Isolation of a recombinant antibody specific for a surface marker of the corneal endothelium by phage display. Scientific Reports 6 : 21661. ScholarBank@NUS Repository. https://doi.org/10.1038/srep21661
Rights: Attribution 4.0 International
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.
Source Title: Scientific Reports
URI: https://scholarbank.nus.edu.sg/handle/10635/178941
ISSN: 20452322
DOI: 10.1038/srep21661
Rights: Attribution 4.0 International
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