Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression

Abstract Using Chlamydia trachomatis (Ct) as a complex model organism, we describe a method to generate bacterial whole-proteome microarrays using cell-free, on-chip protein expression. Expression constructs were generated by two successive PCRs directly from bacterial genomic DNA. Bacterial protein...

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Autores principales:	Katrin Hufnagel, Smiths Lueong, Martina Willhauck-Fleckenstein, Agnes Hotz-Wagenblatt, Beiping Miao, Andrea Bauer, Angelika Michel, Julia Butt, Michael Pawlita, Jörg D. Hoheisel, Tim Waterboer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6e5136bb68724a66a4edce0dbcc7a62e
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spelling	oai:doaj.org-article:6e5136bb68724a66a4edce0dbcc7a62e2021-12-02T11:40:47ZImmunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression10.1038/s41598-018-25918-32045-2322https://doaj.org/article/6e5136bb68724a66a4edce0dbcc7a62e2018-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-25918-3https://doaj.org/toc/2045-2322Abstract Using Chlamydia trachomatis (Ct) as a complex model organism, we describe a method to generate bacterial whole-proteome microarrays using cell-free, on-chip protein expression. Expression constructs were generated by two successive PCRs directly from bacterial genomic DNA. Bacterial proteins expressed on microarrays display antigenic epitopes, thereby providing an efficient method for immunoprofiling of patients and allowing de novo identification of disease-related serum antibodies. Through comparison of antibody reactivity patterns, we newly identified antigens recognized by known Ct-seropositive samples, and antigens reacting only with samples from cervical cancer (CxCa) patients. Large-scale validation experiments using high-throughput suspension bead array serology confirmed their significance as markers for either general Ct infection or CxCa, supporting an association of Ct infection with CxCa. In conclusion, we introduce a method for generation of fast and efficient proteome immunoassays which can be easily adapted for other microorganisms in all areas of infection research.Katrin HufnagelSmiths LueongMartina Willhauck-FleckensteinAgnes Hotz-WagenblattBeiping MiaoAndrea BauerAngelika MichelJulia ButtMichael PawlitaJörg D. HoheiselTim WaterboerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-12 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Katrin Hufnagel Smiths Lueong Martina Willhauck-Fleckenstein Agnes Hotz-Wagenblatt Beiping Miao Andrea Bauer Angelika Michel Julia Butt Michael Pawlita Jörg D. Hoheisel Tim Waterboer Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
description	Abstract Using Chlamydia trachomatis (Ct) as a complex model organism, we describe a method to generate bacterial whole-proteome microarrays using cell-free, on-chip protein expression. Expression constructs were generated by two successive PCRs directly from bacterial genomic DNA. Bacterial proteins expressed on microarrays display antigenic epitopes, thereby providing an efficient method for immunoprofiling of patients and allowing de novo identification of disease-related serum antibodies. Through comparison of antibody reactivity patterns, we newly identified antigens recognized by known Ct-seropositive samples, and antigens reacting only with samples from cervical cancer (CxCa) patients. Large-scale validation experiments using high-throughput suspension bead array serology confirmed their significance as markers for either general Ct infection or CxCa, supporting an association of Ct infection with CxCa. In conclusion, we introduce a method for generation of fast and efficient proteome immunoassays which can be easily adapted for other microorganisms in all areas of infection research.
format	article
author	Katrin Hufnagel Smiths Lueong Martina Willhauck-Fleckenstein Agnes Hotz-Wagenblatt Beiping Miao Andrea Bauer Angelika Michel Julia Butt Michael Pawlita Jörg D. Hoheisel Tim Waterboer
author_facet	Katrin Hufnagel Smiths Lueong Martina Willhauck-Fleckenstein Agnes Hotz-Wagenblatt Beiping Miao Andrea Bauer Angelika Michel Julia Butt Michael Pawlita Jörg D. Hoheisel Tim Waterboer
author_sort	Katrin Hufnagel
title	Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
title_short	Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
title_full	Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
title_fullStr	Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
title_full_unstemmed	Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
title_sort	immunoprofiling of chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/6e5136bb68724a66a4edce0dbcc7a62e
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Immunoprofiling of Chlamydia trachomatis using whole-proteome microarrays generated by on-chip in situ expression

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