Ngubane, NACGresh, LIoerger, TRSacchettini, JCZhang, YJRubin, EJPym, AKhati, M2014-02-132014-02-132013-11Ngubane, N.A.C, Gresh, L, Ioerger, T.R, Sacchettini, J.C, Zhang, Y.J, Rubin, E.J, Pym, A and Khati, M. 2013. High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria. PLOS One, vol. 8(11), pp 1-111932-6203http://www.plosone.org/article/fetchObject.action?uri=info%3Adoi%2F10.1371%2Fjournal.pone.0077844&representation=PDFhttp://hdl.handle.net/10204/7199Copyright: 2013 Public Library of Science. This is an OA journal. The journal authorizes the publication of the information herewith contained. Published in PLOS One, vol. 8(11), pp 1-11Bacterial 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.enMycobacterium tuberculosisM. tbMycobacteriaArticleNgubane, N., Gresh, L., Ioerger, T., Sacchettini, J., Zhang, Y., Rubin, E., ... Khati, M. (2013). High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria. http://hdl.handle.net/10204/7199Ngubane, NAC, L Gresh, TR Ioerger, JC Sacchettini, YJ Zhang, EJ Rubin, A Pym, and M Khati "High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria." (2013) http://hdl.handle.net/10204/7199Ngubane N, Gresh L, Ioerger T, Sacchettini J, Zhang Y, Rubin E, et al. High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria. 2013; http://hdl.handle.net/10204/7199.TY - Article AU - Ngubane, NAC AU - Gresh, L AU - Ioerger, TR AU - Sacchettini, JC AU - Zhang, YJ AU - Rubin, EJ AU - Pym, A AU - Khati, M 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 - 2013-11 DB - ResearchSpace DP - CSIR KW - Mycobacterium tuberculosis KW - M. tb KW - Mycobacteria LK - https://researchspace.csir.co.za PY - 2013 SM - 1932-6203 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/10204/7199 ER -