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

Scientific Reports
Katrin HufnagelTim Waterboer

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.

References

Dec 10, 1999·International Journal of Cancer. Journal International Du Cancer·P KoskelaJ Paavonen
Feb 24, 2001·Cellular Microbiology·T HackstadtE R Fischer
Jun 29, 2004·International Journal of Cancer. Journal International Du Cancer·Jennifer S SmithUNKNOWN IARC multicentric case-control study
Jul 3, 2004·Science·Niroshan RamachandranJoshua LaBaer
Oct 12, 2004·Vaccine·Diane E KawaRichard S Stephens
Jul 27, 2005·FEMS Microbiology Reviews·Yasser M Abdelrahman, Robert J Belland
Jul 11, 2006·Molecular & Cellular Proteomics : MCP·Philipp AngenendtJörg D Hoheisel
Nov 11, 2006·International Journal of Cancer. Journal International Du Cancer·Margaret M MadeleineJanet R Daling
Jun 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Kevin Hybiske, Richard S Stephens
Jan 15, 2008·Current Opinion in Infectious Diseases·Inger J Bakken
Jan 22, 2008·Nature Methods·Mingyue HeMichael J Taussig
Jul 17, 2008·Cancer Epidemiology, Biomarkers & Prevention : a Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology·Bolormaa DondogMichael Pawlita
Jan 7, 2009·BMC Microbiology·Damir T AlzhanovDaniel D Rockey
Jun 29, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Jie WangGuangming Zhong
Apr 20, 2012·International Journal of Cancer. Journal International Du Cancer·Gordana HalecMichael Pawlita
Nov 8, 2012·PLoS Neglected Tropical Diseases·E Brook GoodhewPatrick J Lammie
Feb 14, 2013·Sexually Transmitted Diseases·Catherine Lindsey SatterwhiteHillard Weinstock
Dec 18, 2013·International Journal of Cancer. Journal International Du Cancer·Xavier CastellsaguéElio Riboli
Dec 19, 2013·Archives of Gynecology and Obstetrics·Jani SilvaRui Medeiros
May 13, 2014·The Journal of Infection·Markus SchmittMichael Pawlita
Nov 4, 2015·Journal of Bacteriology·Christopher J Rosario, Ming Tan
Mar 31, 2017·Microbial Cell·Catherine M O'Connell, Morgan E Ferone

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Citations

Apr 30, 2019·Advanced Materials·Daniela S MattesFrank Breitling
Oct 2, 2020·Cancer Epidemiology, Biomarkers & Prevention : a Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology·Rima JeskeKatrin Hufnagel

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Methods Mentioned

BETA
PCRs
ELISA
protein array
PCR
suspension array technology
protein folding

Software Mentioned

SAS
Excel
GenePix Pro
MIST
GraphPad Prism
Perl script

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