Mimtags: the use of phage display technology to produce novel protein-specific probes

Ahmed,N, Dhanapala,P, Sadli,N, Barrow,CJ and Suphioglu,C 2014, Mimtags: the use of phage display technology to produce novel protein-specific probes, Journal of immunological methods, vol. 405, pp. 121-129, doi: 10.1016/j.jim.2014.02.001.

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Title Mimtags: the use of phage display technology to produce novel protein-specific probes
Author(s) Ahmed,N
Barrow,CJORCID iD for Barrow,CJ orcid.org/0000-0002-2153-7267
Suphioglu,CORCID iD for Suphioglu,C orcid.org/0000-0003-0101-0668
Journal name Journal of immunological methods
Volume number 405
Start page 121
End page 129
Total pages 9
Publisher Elsevier
Place of publication Amsterdam, The Netherlands
Publication date 2014-03
ISSN 1872-7905
Keyword(s) Antibodies
Phage display
Bacteriophage M13
Enzyme-Linked Immunosorbent Assay
Epitope Mapping
Peptide Library
Receptors, Interleukin-4
Reproducibility of Results
Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
Summary 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.
Language eng
DOI 10.1016/j.jim.2014.02.001
Field of Research 060113 Synthetic Biology
Socio Economic Objective 920108 Immune System and Allergy
HERDC Research category C1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2014, Elsevier
Persistent URL http://hdl.handle.net/10536/DRO/DU:30071903

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