Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept.
Förster resonance energy transfer (FRET) is a phenomenon widely utilized in biomedical research of macromolecular interactions. In FRET energy is transferred between two fluorophores, the donor and the acceptor. Herein we describe a novel approach utilizing time-resolved FRET (TR-FRET) for the detec...
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2013
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oai:doaj.org-article:7c040746cdfb44dcab90020131d304de2021-11-18T07:46:40ZTime-resolved FRET -based approach for antibody detection - a new serodiagnostic concept.1932-620310.1371/journal.pone.0062739https://doaj.org/article/7c040746cdfb44dcab90020131d304de2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23667515/?tool=EBIhttps://doaj.org/toc/1932-6203Förster resonance energy transfer (FRET) is a phenomenon widely utilized in biomedical research of macromolecular interactions. In FRET energy is transferred between two fluorophores, the donor and the acceptor. Herein we describe a novel approach utilizing time-resolved FRET (TR-FRET) for the detection of antibodies not only in a solution-phase homogenous assay but also in single- and two-step solid-phase assays. Our method is based on the principle that the Y-shaped immunoglobulin G molecule is able to simultaneously bind two identical antigen molecules. Hence, if a specific IgG is mixed with donor- and acceptor-labeled antigens, the binding of antigens can be measured by TR-FRET. Using donor- and acceptor-labeled streptavidins (SAs) in conjunction with a polyclonal and a monoclonal anti-SA antibody we demonstrate that this approach is fully functional. In addition we characterize the immune complexes responsible for the TR-FRET signal using density gradient ultracentrifugation and solid-phase immunoassays. The homogenous TR-FRET assay described provides a rapid and robust tool for antibody detection, with a wide potential in medical diagnostics.Satu SaraheimoJussi HepojokiJussi HepojokiVisa NurmiAnne LahtinenIlkka HemmiläAntti VaheriOlli VapalahtiKlaus HedmanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 5, p e62739 (2013) |
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DOAJ |
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EN |
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Medicine R Science Q |
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Medicine R Science Q Satu Saraheimo Jussi Hepojoki Jussi Hepojoki Visa Nurmi Anne Lahtinen Ilkka Hemmilä Antti Vaheri Olli Vapalahti Klaus Hedman Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept. |
| description |
Förster resonance energy transfer (FRET) is a phenomenon widely utilized in biomedical research of macromolecular interactions. In FRET energy is transferred between two fluorophores, the donor and the acceptor. Herein we describe a novel approach utilizing time-resolved FRET (TR-FRET) for the detection of antibodies not only in a solution-phase homogenous assay but also in single- and two-step solid-phase assays. Our method is based on the principle that the Y-shaped immunoglobulin G molecule is able to simultaneously bind two identical antigen molecules. Hence, if a specific IgG is mixed with donor- and acceptor-labeled antigens, the binding of antigens can be measured by TR-FRET. Using donor- and acceptor-labeled streptavidins (SAs) in conjunction with a polyclonal and a monoclonal anti-SA antibody we demonstrate that this approach is fully functional. In addition we characterize the immune complexes responsible for the TR-FRET signal using density gradient ultracentrifugation and solid-phase immunoassays. The homogenous TR-FRET assay described provides a rapid and robust tool for antibody detection, with a wide potential in medical diagnostics. |
| format |
article |
| author |
Satu Saraheimo Jussi Hepojoki Jussi Hepojoki Visa Nurmi Anne Lahtinen Ilkka Hemmilä Antti Vaheri Olli Vapalahti Klaus Hedman |
| author_facet |
Satu Saraheimo Jussi Hepojoki Jussi Hepojoki Visa Nurmi Anne Lahtinen Ilkka Hemmilä Antti Vaheri Olli Vapalahti Klaus Hedman |
| author_sort |
Satu Saraheimo |
| title |
Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept. |
| title_short |
Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept. |
| title_full |
Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept. |
| title_fullStr |
Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept. |
| title_full_unstemmed |
Time-resolved FRET -based approach for antibody detection - a new serodiagnostic concept. |
| title_sort |
time-resolved fret -based approach for antibody detection - a new serodiagnostic concept. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2013 |
| url |
https://doaj.org/article/7c040746cdfb44dcab90020131d304de |
| work_keys_str_mv |
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| _version_ |
1718422981751865344 |