Abstract
The detection of weakly expressed proteins and protein complexes in biological samples represents a fundamental challenge. We have developed a new proximity-ligation strategy named 3PLA that uses three recognition events for the highly specific and sensitive detection of as little as a hundred molecules of the vascular endothelial growth factor (VEGF), the biomarkers troponin I, and prostate-specific antigen (PSA) alone or in complex with an inhibitor—demonstrating the versatility of 3PLA.
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Acknowledgements
This work was supported by the EU-FP6 integrated project MolTools, the Swedish Research Councils for Medicine and for Natural and Engineering Sciences, the Wallenberg Consortium North for Functional Genomics, and the Uppsala Bio-X.
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Contributions
E.S. and U.L. conceived the described method. E.S. designed the experiments, and E.S., E.O. and L.S. performed laboratory work. S.E. and K.P. contributed with the troponin application, U.-H.S. contributed with the PSA application and O.E. contributed to the technical development. E.S. and U.L. wrote the manuscript, and all authors contributed to subsequent drafts.
Note: Supplementary information is available on the Nature Methods website.
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Competing interests
E.S. and U.L. are inventors of techniques used in this publication. U.L. is the founder of Olink AB, which has the rights to these techniques.
Supplementary information
Supplementary Table 1
3PLA oligonucleotide sequences.
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Schallmeiner, E., Oksanen, E., Ericsson, O. et al. Sensitive protein detection via triple-binder proximity ligation assays. Nat Methods 4, 135–137 (2007). https://doi.org/10.1038/nmeth974
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DOI: https://doi.org/10.1038/nmeth974
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