Please use this identifier to cite or link to this item: https://doi.org/10.3390/ijms13044937
Title: Generation and characterization of a novel recombinant antibody against 15-ketocholestane isolated by phage-display
Authors: Omedul Islam M. 
Lim Y.T. 
Chan C.E.Z. 
Cazenave-Gassiot A. 
Ludovic Croxford J. 
Wenk M.R. 
Macary P.A. 
Hanson B.J. 
Keywords: antigen
biological marker
immunoglobulin G antibody
monoclonal antibody
monoclonal antibody 2E9
oxysterol
oxysterol 15 ketocholestane
recombinant oxysterol 15 ketocholestane monoclonal antibody
unclassified drug
biological marker
cholest 4 en 3 one
cholest-8(14)-en-3-ol-15-one
cholesterol
monoclonal antibody
peptide library
allergic encephalomyelitis
animal model
antibody detection
antibody isolation
antibody production
antibody specificity
antigen binding
article
binding affinity
controlled study
disease marker
feasibility study
human
immunogenicity
mass spectrometry
mouse
nonhuman
phage display
analogs and derivatives
animal
blood
cell surface display
diagnostic use
Encephalomyelitis, Autoimmune, Experimental
immunology
multiple sclerosis
peptide library
Animals
Antibodies, Monoclonal
Biological Markers
Cell Surface Display Techniques
Cholestenones
Cholesterol
Encephalomyelitis, Autoimmune, Experimental
Humans
Mice
Multiple Sclerosis
Peptide Library
Issue Date: 2012
Citation: Omedul Islam M., Lim Y.T., Chan C.E.Z., Cazenave-Gassiot A., Ludovic Croxford J., Wenk M.R., Macary P.A., Hanson B.J. (2012). Generation and characterization of a novel recombinant antibody against 15-ketocholestane isolated by phage-display. International Journal of Molecular Sciences 13 (4) : 4937-4948. ScholarBank@NUS Repository. https://doi.org/10.3390/ijms13044937
Abstract: The employment of monoclonal antibodies (Mabs) to identify disease-associated biomarkers in clinical samples represents the underlying principle for many diagnostic tests. To date, these have been principally developed for protein targets with few reported applications for lipids due to their hydrophobicity and poor immunogenicity. Oxysterols represent a family of lipids implicated in diverse human diseases where Mab-based detection assays could have a profound effect on their utility as clinical biomarkers. These are usually identified in patients' samples by mass- spectrometry based approaches. Here, we describe an antibody phage-library based screening methodology for generating a recombinant monoclonal antibody (RAb) targeting the oxysterol-15-ketocholestane (15-KA), a lipid implicated in multiple sclerosis and Autoimmune Encephalomyelitis (EAE). The antibody is highly specific for 15-KA and shows little or no binding activity for other closely related oxysterols. We employ RAb2E9 to address the controversy over whether 15-KA is a true biomarker for MS/EAE and show that 15-KA is undetectable in serum taken from mice with EAE using antibody based detection methodologies; a finding confirmed by mass-spectrometry analysis. This study demonstrates the technical feasibility of using phage display to isolate highly specific antibodies against poorly immunogenic, small molecule lipids. © 2012 by the authors.
Source Title: International Journal of Molecular Sciences
URI: https://scholarbank.nus.edu.sg/handle/10635/174658
ISSN: 1422-0067
DOI: 10.3390/ijms13044937
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