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Mimtags: the use of phage display technology to produce novel protein-specific probes
journal contribution
posted on 2014-03-01, 00:00 authored by Nayyar Ahmed, Pathum Dhanapala, Nadia Sadli, Colin BarrowColin Barrow, Cenk SuphiogluCenk SuphiogluIn recent times the use of protein-specific probes in the field of proteomics has undergone evolutionary changes leading to the discovery of new probing techniques. Protein-specific probes serve two main purposes: epitope mapping and detection assays. One such technique is the use of phage display in the random selection of peptide mimotopes (mimtags) that can tag epitopes of proteins, replacing the use of monoclonal antibodies in detection systems. In this study, phage display technology was used to screen a random peptide library with a biologically active purified human interleukin-4 receptor (IL-4R) and interleukin-13 (IL-13) to identify mimtag candidates that interacted with these proteins. Once identified, the mimtags were commercially synthesised, biotinylated and used for in vitro immunoassays. We have used phage display to identify M13 phage clones that demonstrated specific binding to IL-4R and IL-13 cytokine. A consensus in binding sequences was observed and phage clones characterised had identical peptide sequence motifs. Only one was synthesised for use in further immunoassays, demonstrating significant binding to either IL-4R or IL-13. We have successfully shown the use of phage display to identify and characterise mimtags that specifically bind to their target epitope. Thus, this new method of probing proteins can be used in the future as a novel tool for immunoassay and detection technique, which is cheaper and more rapidly produced and therefore a better alternative to the use of monoclonal antibodies.
History
Journal
Journal of immunological methodsVolume
405Pagination
121 - 129Publisher
ElsevierLocation
Amsterdam, The NetherlandsPublisher DOI
eISSN
1872-7905Language
engPublication classification
C Journal article; C1 Refereed article in a scholarly journalCopyright notice
2014, ElsevierUsage metrics
Categories
Keywords
AntibodiesEpitopeMimotopesMimtagsPhage displayProbesBacteriophage M13Enzyme-Linked Immunosorbent AssayEpitope MappingEpitopesHumansInterleukin-13Peptide LibraryPeptidesReceptors, Interleukin-4Reproducibility of ResultsScience & TechnologyLife Sciences & BiomedicineBiochemical Research MethodsImmunologyBiochemistry & Molecular BiologyMONOCLONAL-ANTIBODIESPEPTIDE LIBRARIESIGE-EPITOPESMIMOTOPEALLERGENDIAGNOSTICSAFFINITYDEFINEImmunology
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