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Bioluminescent sensor proteins for point-of-care therapeutic drug monitoring


For many drugs, finding the balance between efficacy and toxicity requires monitoring their concentrations in the patient's blood. Quantifying drug levels at the bedside or at home would have advantages in terms of therapeutic outcome and convenience, but current techniques require the setting of a diagnostic laboratory. We have developed semisynthetic bioluminescent sensors that permit precise measurements of drug concentrations in patient samples by spotting minimal volumes on paper and recording the signal using a simple point-and-shoot camera. Our sensors have a modular design consisting of a protein-based and a synthetic part and can be engineered to selectively recognize a wide range of drugs, including immunosuppressants, antiepileptics, anticancer agents and antiarrhythmics. This low-cost point-of-care method could make therapies safer, increase the convenience of doctors and patients and make therapeutic drug monitoring available in regions with poor infrastructure.

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Figure 1: Design and performance of a sensor for methotrexate.
Figure 2: Measuring methotrexate concentrations in patient samples using a point-and-shoot digital camera.
Figure 3: LUCID for tacrolimus and sirolimus.
Figure 4: LUCID for cyclosporin A.
Figure 5: LUCIDs for topiramate and digoxin.

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  1. Saint-Marcoux, F., Sauvage, F.L. & Marquet, P. Current role of LC-MS in therapeutic drug monitoring. Anal. Bioanal. Chem. 388, 1327–1349 (2007).

    Article  CAS  Google Scholar 

  2. Brun, M.A., Tan, K.T., Nakata, E., Hinner, M.J. & Johnsson, K. Semisynthetic fluorescent sensor proteins based on self-labeling protein tags. J. Am. Chem. Soc. 131, 5873–5884 (2009).

    Article  CAS  Google Scholar 

  3. Brun, M.A. et al. A semisynthetic fluorescent sensor protein for glutamate. J. Am. Chem. Soc. 134, 7676–7678 (2012).

    Article  CAS  Google Scholar 

  4. Brun, M.A. et al. Semisynthesis of fluorescent metabolite sensors on cell surfaces. J. Am. Chem. Soc. 133, 16235–16242 (2011).

    Article  CAS  Google Scholar 

  5. Masharina, A., Reymond, L., Maurel, D., Umezawa, K. & Johnsson, K. A fluorescent sensor for GABA and synthetic GABAB receptor ligands. J. Am. Chem. Soc. 134, 19026–19034 (2012).

    Article  CAS  Google Scholar 

  6. Lennard, L. Therapeutic drug monitoring of cytotoxic drugs. Br. J. Clin. Pharmacol. 52 (suppl. 1): 75S–87S (2001).

    Article  CAS  Google Scholar 

  7. Keppler, A. et al. A general method for the covalent labeling of fusion proteins with small molecules in vivo. Nat. Biotechnol. 21, 86–89 (2003).

    Article  CAS  Google Scholar 

  8. Saito, K. et al. Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level. PLoS ONE 5, e9935 (2010).

    Article  Google Scholar 

  9. Iwakura, M. & Nakamura, T. Effects of the length of a glycine linker connecting the N- and C-termini of a circularly permuted dihydrofolate reductase. Protein Eng. 11, 707–713 (1998).

    Article  CAS  Google Scholar 

  10. Jaffe, N. & Gorlick, R. High-dose methotrexate in osteosarcoma: let the questions surcease—time for final acceptance. J. Clin. Oncol. 26, 4365–4366 (2008).

    Article  CAS  Google Scholar 

  11. Al-Turkmani, M.R., Law, T., Narla, A. & Kellogg, M.D. Difficulty measuring methotrexate in a patient with high-dose methotrexate-induced nephrotoxicity. Clin. Chem. 56, 1792–1794 (2010).

    Article  CAS  Google Scholar 

  12. Kratz, A., Ferraro, M., Sluss, P.M. & Lewandrowski, K.B. Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Laboratory reference values. N. Engl. J. Med. 351, 1548–1563 (2004); erratum 351, 2461.

    Article  CAS  Google Scholar 

  13. Pollock, N.R. et al. A paper-based multiplexed transaminase test for low-cost, point-of-care liver function testing. Sci. Transl. Med. 4, 152ra129 (2012).

    Article  Google Scholar 

  14. Armstrong, V.W. & Oellerich, M. New developments in the immunosuppressive drug monitoring of cyclosporine, tacrolimus, and azathioprine. Clin. Biochem. 34, 9–16 (2001).

    Article  CAS  Google Scholar 

  15. Röhrig, C.H., Loch, C., Guan, J.Y., Siegal, G. & Overhand, M. Fragment-based synthesis and SAR of modified FKBP ligands: influence of different linking on binding affinity. ChemMedChem 2, 1054–1070 (2007).

    Article  Google Scholar 

  16. Patsalos, P.N. et al. Antiepileptic drugs—best practice guidelines for therapeutic drug monitoring: a position paper by the subcommission on therapeutic drug monitoring, ILAE Commission on Therapeutic Strategies. Epilepsia 49, 1239–1276 (2008).

    Article  CAS  Google Scholar 

  17. Valdes, R. Jr., Jortani, S.A. & Gheorghiade, M. Standards of laboratory practice: cardiac drug monitoring. National Academy of Clinical Biochemistry. Clin. Chem. 44, 1096–1109 (1998).

    CAS  PubMed  Google Scholar 

  18. Matsui, H. & Schwartz, A. Mechanism of cardiac glycoside inhibition of the (Na+-K+)-dependent ATPase from cardiac tissue. Biochim. Biophys. Acta 151, 655–663 (1968).

    Article  CAS  Google Scholar 

  19. Tinberg, C.E. et al. Computational design of ligand-binding proteins with high affinity and selectivity. Nature 501, 212–216 (2013).

    Article  CAS  Google Scholar 

  20. Terra, S.G., Washam, J.B., Dunham, G.D. & Gattis, W.A. Therapeutic range of digoxin's efficacy in heart failure: what is the evidence? Pharmacotherapy 19, 1123–1126 (1999).

    Article  CAS  Google Scholar 

  21. Smith, T.W. & Haber, E. Digoxin intoxication: the relationship of clinical presentation to serum digoxin concentration. J. Clin. Invest. 49, 2377–2386 (1970).

    Article  CAS  Google Scholar 

  22. Turner, A.P. Biosensors: sense and sensibility. Chem. Soc. Rev. 42, 3184–3196 (2013).

    Article  CAS  Google Scholar 

  23. Krishnamurthy, V.M., Semetey, V., Bracher, P.J., Shen, N. & Whitesides, G.M. Dependence of effective molarity on linker length for an intramolecular protein-ligand system. J. Am. Chem. Soc. 129, 1312–1320 (2007).

    Article  CAS  Google Scholar 

  24. Sawaya, M.R. & Kraut, J. Loop and subdomain movements in the mechanism of Escherichia coli dihydrofolate reductase: crystallographic evidence. Biochemistry 36, 586–603 (1997).

    Article  CAS  Google Scholar 

  25. Dubowchik, G.M. et al. 2-Aryl-2,2-difluoroacetamide FKBP12 ligands: synthesis and X-ray structural studies. Org. Lett. 3, 3987–3990 (2001).

    Article  CAS  Google Scholar 

  26. Schulz, M. & Schmoldt, A. Therapeutic and toxic blood concentrations of more than 800 drugs and other xenobiotics. Pharmazie 58, 447–474 (2003).

    CAS  Google Scholar 

  27. Srivastava, D.K. et al. Structural analysis of charge discrimination in the binding of inhibitors to human carbonic anhydrases I and II. J. Am. Chem. Soc. 129, 5528–5537 (2007).

    Article  CAS  Google Scholar 

  28. Schneider, C.A., Rasband, W.S. & Eliceiri, K.W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).

    Article  CAS  Google Scholar 

  29. Li, C.H. & Tam, P.K.S. An iterative algorithm for minimum cross entropy thresholding. Pattern Recognit. Lett. 19, 771–776 (1998).

    Article  Google Scholar 

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This work was supported by École Polytechnique Fédérale de Lausanne, the Swiss National Science Foundation, the National Centre of Competence in Research Chemical Biology, and the Defense Threat Reduction Agency. We are grateful to T. Buclin, N. Widmer and L. Decosterd from the CHUV for helpful discussions.

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Authors and Affiliations



R.G., A.S. and K.J. designed experiments, R.G. and A.S. performed experiments, A.S. and L.R. performed chemical synthesis and characterization of the sensor ligands, L.P. wrote software for image analysis, D.W. provided patient samples, and C.E.T. and D.B. provided DIG10.3. All of the authors contributed to the writing of the manuscript.

Corresponding author

Correspondence to Kai Johnsson.

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Competing interests

R.G., A.S. and K.J. have filed a Patent Cooperation Treaty (World Intellectual Property Organization) patent application on the design and use of LUCIDs.

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Supplementary Notes 1 and 2 and Supplementary Results. (PDF 4186 kb)

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Griss, R., Schena, A., Reymond, L. et al. Bioluminescent sensor proteins for point-of-care therapeutic drug monitoring. Nat Chem Biol 10, 598–603 (2014).

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