High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria

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dc.contributor.authorNgubane, Nqobile A Cen_ZA
dc.contributor.authorGresh, Lionelen_ZA
dc.contributor.authorIoerger, Thomas Ren_ZA
dc.contributor.authorSacchettini, James Cen_ZA
dc.contributor.authorZhang, Yanjia Jen_ZA
dc.contributor.authorRubin, Eric Jen_ZA
dc.contributor.authorPym, Alexanderen_ZA
dc.contributor.authorKhati, Makobetsaen_ZA
dc.date.accessioned2015-11-23T12:36:53Z
dc.date.available2015-11-23T12:36:53Z
dc.date.issued2013en_ZA
dc.description.abstractBacterial cell wall components have been previously used as infection biomarkers detectable by antibodies. However, it is possible that the surface of the Mycobacterium tuberculosis ( M. tb ), the causative agent of tuberculosis (TB), also possesses molecules which might be non-antigenic. This makes the probing of biomarkers on the surface of M. tb cell wall difficult using antibodies. Here we demonstrate the use of phage display technology to identify peptides that bind to mycobacteria. We identified these clones using both random clone picking and high throughput sequencing. We demonstrate that random clone picking does not necessarily identify highly enriched clones. We further showed that the clone displaying the CPLHARLPC peptide which was identified by Illumina sequencing as the most enriched, binds better to mycobacteria than three clones selected by random picking. Using surface plasmon resonance, we showed that chemically synthesised CPLHARLPC peptide binds to a 15 KDa peptide from M.tb H37Rv whole cell lysates. These observations demonstrate that phage display technology combined with high-throughput sequencing is a powerful tool to identify peptides that can be used for investigating potential non-antigenic biomarkers for TB and other bacterial infections.en_ZA
dc.identifier.apacitationNgubane, N. A. C., Gresh, L., Ioerger, T. R., Sacchettini, J. C., Zhang, Y. J., Rubin, E. J., ... Khati, M. (2013). High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria. <i>PLoS One</i>, http://hdl.handle.net/11427/15343en_ZA
dc.identifier.chicagocitationNgubane, Nqobile A C, Lionel Gresh, Thomas R Ioerger, James C Sacchettini, Yanjia J Zhang, Eric J Rubin, Alexander Pym, and Makobetsa Khati "High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria." <i>PLoS One</i> (2013) http://hdl.handle.net/11427/15343en_ZA
dc.identifier.citationNgubane, N. A., Gresh, L., Ioerger, T. R., Sacchettini, J. C., Zhang, Y. J., Rubin, E. J., ... & Khati, M. (2012). High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria. PloS one, 8(11), e77844-e77844. doi:10.1371/journal.pone.0077844en_ZA
dc.identifier.ris TY - Journal Article AU - Ngubane, Nqobile A C AU - Gresh, Lionel AU - Ioerger, Thomas R AU - Sacchettini, James C AU - Zhang, Yanjia J AU - Rubin, Eric J AU - Pym, Alexander AU - Khati, Makobetsa AB - Bacterial cell wall components have been previously used as infection biomarkers detectable by antibodies. However, it is possible that the surface of the Mycobacterium tuberculosis ( M. tb ), the causative agent of tuberculosis (TB), also possesses molecules which might be non-antigenic. This makes the probing of biomarkers on the surface of M. tb cell wall difficult using antibodies. Here we demonstrate the use of phage display technology to identify peptides that bind to mycobacteria. We identified these clones using both random clone picking and high throughput sequencing. We demonstrate that random clone picking does not necessarily identify highly enriched clones. We further showed that the clone displaying the CPLHARLPC peptide which was identified by Illumina sequencing as the most enriched, binds better to mycobacteria than three clones selected by random picking. Using surface plasmon resonance, we showed that chemically synthesised CPLHARLPC peptide binds to a 15 KDa peptide from M.tb H37Rv whole cell lysates. These observations demonstrate that phage display technology combined with high-throughput sequencing is a powerful tool to identify peptides that can be used for investigating potential non-antigenic biomarkers for TB and other bacterial infections. DA - 2013 DB - OpenUCT DO - 10.1371/journal.pone.0077844 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2013 T1 - High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria TI - High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria UR - http://hdl.handle.net/11427/15343 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/15343
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0077844
dc.identifier.vancouvercitationNgubane NAC, Gresh L, Ioerger TR, Sacchettini JC, Zhang YJ, Rubin EJ, et al. High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria. PLoS One. 2013; http://hdl.handle.net/11427/15343.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentDepartment of Medicineen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_ZA
dc.rights.holder© 2013 Ngubane et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherMycobacterium tuberculosisen_ZA
dc.subject.otherPhage displayen_ZA
dc.subject.otherCloningen_ZA
dc.subject.otherPeptide librariesen_ZA
dc.subject.otherBacteriophagesen_ZA
dc.subject.otherTuberculosisen_ZA
dc.subject.otherBovine tuberculosisen_ZA
dc.subject.otherMycobacteriaen_ZA
dc.titleHigh-throughput sequencing enhanced phage display identifies peptides that bind mycobacteriaen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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