Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.

Campylobacter jejuni remains one of the major gut pathogens of our time. Its zoonotic nature and wide-spread distribution in industrialized countries calls for a quick and reliable diagnostic tool. Antibody-based detection presents a suitable means to identify pathogenic bacteria. However, the knowl...

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Autores principales:	Sebastian Hoppe, Frank F Bier, Markus von Nickisch-Rosenegk
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/057b2857539e481c94beabfa9acb693f
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spelling	oai:doaj.org-article:057b2857539e481c94beabfa9acb693f2021-11-18T07:43:51ZRapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.1932-620310.1371/journal.pone.0065837https://doaj.org/article/057b2857539e481c94beabfa9acb693f2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23734261/?tool=EBIhttps://doaj.org/toc/1932-6203Campylobacter jejuni remains one of the major gut pathogens of our time. Its zoonotic nature and wide-spread distribution in industrialized countries calls for a quick and reliable diagnostic tool. Antibody-based detection presents a suitable means to identify pathogenic bacteria. However, the knowledge about immunodominant targets is limited. Thus, an approach is presented, which allows for the rapid screening of numerous cDNA derived expression clones to identify novel antigens. The deeper understanding of immunodominant proteins assists in the design of diagnostic tools and furthers the insight into the bacterium's pathogenicity as well as revealing potential candidates for vaccination. We have successfully screened 1536 clones of an expression library to identify 22 proteins that have not been described as immunodominant before. After subcloning the corresponding 22 genes and expression of full-length proteins, we investigated the immunodominant character by microarrays and ELISA. Subsequently, seven proteins were selected for epitope mapping. For cj0669 and cj0920c linear epitopes were identified. For cj0669, specificity assays revealed a specific linear epitope site. Consequently, an eleven amino acid residue sequence TLIKELKRLGI was analyzed via alanine scan, which revealed the glycine residue to be significant for binding of the antibody. The innovative approach presented herein of generating cDNAs of prokaryotes in combination with a microarray platform rendering time-consuming purification steps obsolete has helped to illuminate novel immunodominant proteins of C.jejuni. The findings of a specific linear epitope pave the way for a plethora of future research and the potential use in diagnostic applications such as serological screenings. Moreover, the current approach is easily adaptable to other highly relevant bacteria making it a formidable tool for the future discovery of antigens and potential biomarkers. Consequently, it is desirable to simplify the identification of structural epitopes, as this would extend the spectrum of novel epitopes to be detected.Sebastian HoppeFrank F BierMarkus von Nickisch-RosenegkPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 5, p e65837 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sebastian Hoppe Frank F Bier Markus von Nickisch-Rosenegk Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.
description	Campylobacter jejuni remains one of the major gut pathogens of our time. Its zoonotic nature and wide-spread distribution in industrialized countries calls for a quick and reliable diagnostic tool. Antibody-based detection presents a suitable means to identify pathogenic bacteria. However, the knowledge about immunodominant targets is limited. Thus, an approach is presented, which allows for the rapid screening of numerous cDNA derived expression clones to identify novel antigens. The deeper understanding of immunodominant proteins assists in the design of diagnostic tools and furthers the insight into the bacterium's pathogenicity as well as revealing potential candidates for vaccination. We have successfully screened 1536 clones of an expression library to identify 22 proteins that have not been described as immunodominant before. After subcloning the corresponding 22 genes and expression of full-length proteins, we investigated the immunodominant character by microarrays and ELISA. Subsequently, seven proteins were selected for epitope mapping. For cj0669 and cj0920c linear epitopes were identified. For cj0669, specificity assays revealed a specific linear epitope site. Consequently, an eleven amino acid residue sequence TLIKELKRLGI was analyzed via alanine scan, which revealed the glycine residue to be significant for binding of the antibody. The innovative approach presented herein of generating cDNAs of prokaryotes in combination with a microarray platform rendering time-consuming purification steps obsolete has helped to illuminate novel immunodominant proteins of C.jejuni. The findings of a specific linear epitope pave the way for a plethora of future research and the potential use in diagnostic applications such as serological screenings. Moreover, the current approach is easily adaptable to other highly relevant bacteria making it a formidable tool for the future discovery of antigens and potential biomarkers. Consequently, it is desirable to simplify the identification of structural epitopes, as this would extend the spectrum of novel epitopes to be detected.
format	article
author	Sebastian Hoppe Frank F Bier Markus von Nickisch-Rosenegk
author_facet	Sebastian Hoppe Frank F Bier Markus von Nickisch-Rosenegk
author_sort	Sebastian Hoppe
title	Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.
title_short	Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.
title_full	Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.
title_fullStr	Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.
title_full_unstemmed	Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.
title_sort	rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of campylobacter jejuni.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/057b2857539e481c94beabfa9acb693f
work_keys_str_mv	AT sebastianhoppe rapididentificationofnovelimmunodominantproteinsandcharacterizationofaspecificlinearepitopeofcampylobacterjejuni AT frankfbier rapididentificationofnovelimmunodominantproteinsandcharacterizationofaspecificlinearepitopeofcampylobacterjejuni AT markusvonnickischrosenegk rapididentificationofnovelimmunodominantproteinsandcharacterizationofaspecificlinearepitopeofcampylobacterjejuni
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Rapid identification of novel immunodominant proteins and characterization of a specific linear epitope of Campylobacter jejuni.

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