title: Mimtags: the use of phage display technology to produce novel protein-specific probes.
abstract: In 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.
authors: NeuroAllergy Research Laboratory (NARL), Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216, Australia; School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216 Australia.
authors: Nayyar
authors: N
authors: Ahmed
authors: NeuroAllergy Research Laboratory (NARL), Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216, Australia; School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216 Australia.
authors: Pathum
authors: P
authors: Dhanapala
authors: NeuroAllergy Research Laboratory (NARL), Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216, Australia; School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216 Australia.
authors: Nadia
authors: N
authors: Sadli
authors: School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216 Australia.
authors: Colin J
authors: CJ
authors: Barrow
authors: NeuroAllergy Research Laboratory (NARL), Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216, Australia; School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216 Australia. Electronic address: cenk.suphioglu@deakin.edu.au.
authors: Cenk
authors: C
authors: Suphioglu
mesh_terms: D017104:Bacteriophage M13 / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D018793:Interleukin-13 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019948:Receptors, Interleukin-4 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D000939:Epitopes; D018793:Interleukin-13; D019151:Peptide Library; D010455:Peptides; D019948:Receptors, Interleukin-4
keywords: Antibodies; Epitope; Mimotopes; Mimtags; Phage display; Probes