Next Article in Journal
A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex
Previous Article in Journal
Automatic Extraction of Optimal Endmembers from Airborne Hyperspectral Imagery Using Iterative Error Analysis (IEA) and Spectral Discrimination Measurements
Previous Article in Special Issue
Immobilization Techniques for Microarray: Challenges and Applications
Article Menu

Export Article

Open AccessArticle
Sensors 2015, 15(2), 2614-2628; doi:10.3390/s150202614

Regeneration of Recombinant Antigen Microarrays for the Automated Monitoring of Antibodies against Zoonotic Pathogens in Swine Sera

Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universität München, Marchioninistrasse 17, 81377 Munich, Germany
*
Author to whom correspondence should be addressed.
Received: 31 October 2014 / Accepted: 19 January 2015 / Published: 23 January 2015
(This article belongs to the Special Issue Microarray Sensors)
View Full-Text   |   Download PDF [1595 KB, uploaded 23 January 2015]   |  

Abstract

The ability to regenerate immobilized proteins like recombinant antigens (rAgs) on surfaces is an unsolved problem for flow-based immunoassays on microarray analysis systems. The regeneration on microarray chip surfaces is achieved by changing the protein structures and desorption of antibodies. Afterwards, reactivation of immobilized protein antigens is necessary for reconstitution processes. Any backfolding should be managed in a way that antibodies are able to detect the protein antigens in the next measurement cycle. The regeneration of rAg microarrays was examined for the first time on the MCR3 flow-based chemiluminescence (CL) microarray analysis platform. The aim was to reuse rAg microarray chips in order to reduce the screening effort and costs. An antibody capturing format was used to detect antibodies against zoonotic pathogens in sera of slaughtered pigs. Different denaturation and reactivation buffers were tested. Acidic glycine-SDS buffer (pH 2.5) and 8 M guanidinium hydrochloride showed the best results in respect of denaturation efficiencies. The highest CL signals after regeneration were achieved with a carbonate buffer containing 10 mM DTT and 0.1% BSA for reactivation. Antibodies against Yersinia spp. and hepatitis E virus (HEV) were detected in swine sera on one immunochip over 4 days and 25 measurement cycles. Each cycle took 10 min for detection and regeneration. By using the rAg microarray chip, a fast and automated screening of antibodies against pathogens in sera of slaughtered pigs would be possible for zoonosis monitoring. View Full-Text
Keywords: chemiluminescence microarrays; indirect microarray immunoassays; regeneration; automated analysis platforms; food safety; zoonotic pathogens; recombinant antigen; meat processing; serology chemiluminescence microarrays; indirect microarray immunoassays; regeneration; automated analysis platforms; food safety; zoonotic pathogens; recombinant antigen; meat processing; serology
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Meyer, V.K.; Kober, C.; Niessner, R.; Seidel, M. Regeneration of Recombinant Antigen Microarrays for the Automated Monitoring of Antibodies against Zoonotic Pathogens in Swine Sera. Sensors 2015, 15, 2614-2628.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Sensors EISSN 1424-8220 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top