Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Exploiting NanoLuc luciferase for smartphone-based bioluminescence cell biosensor for (anti)-inflammatory activity and toxicity


The availability of smartphones with high-performance digital image sensors and processing power has completely reshaped the landscape of point-of-need analysis. Thanks to the high maturity level of reporter gene technology and the availability of several bioluminescent proteins with improved features, we were able to develop a bioluminescence smartphone-based biosensing platform exploiting the highly sensitive NanoLuc luciferase as reporter. A 3D-printed smartphone-integrated cell biosensor based on genetically engineered Hek293T cells was developed. Quantitative assessment of (anti)-inflammatory activity and toxicity of liquid samples was performed with a simple and rapid add-and-measure procedure. White grape pomace extracts, known to contain several bioactive compounds, were analyzed, confirming the suitability of the smartphone biosensing platform for analysis of untreated complex biological matrices. Such approach could meet the needs of small medium enterprises lacking fully equipped laboratories for first-level safety tests and rapid screening of new bioactive products.

Smartphone-based bioluminescence cell biosensor

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. 1.

    Spethmann S, Prescher S, Dreger H, Nettlau H, Baumann G, Knebel F, et al. Electrocardiographic monitoring during marathon running: a proof of feasibility for a new telemedical approach. Eur J Prev Cardiol. 2014;21:32–7.

  2. 2.

    Garabedian LF, Ross-Degnan D, Wharam JF. Mobile phone and smartphone technologies for diabetes care and self-management. Curr Diab Rep. 2015;15:109.

  3. 3.

    Roda A, Guardigli M, Calabria D, Calabretta MM, Cevenini L, Michelini E. A 3D-printed device for a smartphone-based chemiluminescence biosensor for lactate in oral fluid and sweat. Analyst. 2014;139:6494–501.

  4. 4.

    Zhang D, Jiang J, Chen J, Zhang Q, Lu Y, Yao Y, et al. Smartphone-based portable biosensing system using impedance measurement with printed electrodes for 2,4,6-trinitrotoluene (TNT) detection. Biosens Bioelectron. 2015;70:81–8.

  5. 5.

    Yin K, Zhang W, Chen L. Pyoverdine secreted by Pseudomonas aeruginosa as a biological recognition element for the fluorescent detection of furazolidone. Biosens Bioelectron. 2014;51:90–6.

  6. 6.

    Li B, Longwen F, Zhang W, Feng W, Chen L. Portable paper-based device for quantitative colorimetric assays relying on light reflectance principle. Electrophoresis. 2014;35:1152–9.

  7. 7.

    Turner AP. Biosensors: sense and sensibility. Chem Soc Rev. 2013;42:3184–96.

  8. 8.

    Comina G, Suska A, Filippini D. Towards autonomous lab-on-a-chip devices for cell phone biosensing. Biosens Bioelectron. 2016;77:1153–67.

  9. 9.

    Mirasoli M, Nascetti A, Caputo D, Zangheri M, Scipinotti R, Cevenini L, et al. Multiwell cartridge with integrated array of amorphous silicon photosensors for chemiluminescence detection: development, characterization and comparison with cooled-CCD luminograph. Anal Bioanal Chem. 2014;406:5645–56.

  10. 10.

    Roda A, Cevenini L, Borg S, Michelini E, Calabretta MM, Schüler D. Bioengineered bioluminescent magnetotactic bacteria as a powerful tool for chip-based whole-cell biosensors. Lab Chip. 2013;13:4881–9.

  11. 11.

    Roda A, Cevenini L, Michelini E, Branchini BR. A portable bioluminescence engineered cell-based biosensor for on-site applications. Biosens Bioelectron. 2011;26:3647–53.

  12. 12.

    Zhang D, Liu Q. Biosensors and bioelectronics on smartphone for portable biochemical detection. Biosens Bioelectron. 2016;75:273–84.

  13. 13.

    Roda A, Mirasoli M, Michelini E, Di Fusco M, Zangheri M, Cevenini L, et al. Progress in chemical luminescence-based biosensors: a critical review. Biosens Bioelectron. 2016;76:164–79.

  14. 14.

    Liu X, Lin TY, Lillehoj PB. Smartphones for cell and biomolecular detection. Ann Biomed Eng. 2014;42:2205–17.

  15. 15.

    Vashist SK, Mudanyali O, Schneider EM, Zengerle R, Ozcan A. Cellphone-based devices for bioanalytical sciences. Anal Bioanal Chem. 2014;406:3263–77.

  16. 16.

    Liao SC, Peng J, Mauk MG, Awasthi S, Song J, Friedman H, et al. Smart cup: a minimally-instrumented, smartphone-based point-of-care molecular diagnostic device. Sens Actuators B Chem. 2016;229:232–38.

  17. 17.

    Petryayeva E, Algar WR. Multiplexed homogeneous assays of proteolytic activity using a smartphone and quantum dots. Anal Chem. 2014;86:3195–202.

  18. 18.

    Nicolini AM, Fronczek CF, Yoon JY. Droplet-based immunoassay on a ‘sticky’ nanofibrous surface for multiplexed and dual detection of bacteria using smartphones. Biosens Bioelectron. 2015;67:560–9.

  19. 19.

    Cevenini L, Calabretta MM, Tarantino G, Michelini E, Roda A. Smartphone-interfaced 3D printed toxicity biosensor integrating bioluminescent “sentinel cells”. Sens Actuators B: Chem. 2016;225:249–57.

  20. 20.

    Charrier T, Durand MJ, Jouanneau S, Dion M, Pernetti M, Poncelet D, et al. A multi-channel bioluminescent bacterial biosensor for the on-line detection of metals and toxicity. Part I: design and optimization of bioluminescent bacterial strains. Anal Bioanal Chem. 2011;400:1051–60.

  21. 21.

    Bazin I, Seo HB, Suehs CM, Ramuz M, De Waard M, Gu MB. Profiling the biological effects of wastewater samples via bioluminescent bacterial biosensors combined with estrogenic assays. Environ Sci Pollut Res Int. 2016. doi:10.1007/s11356-016-6050-5.

  22. 22.

    Elad T, Seo HB, Belkin S, Gu MB. High-throughput prescreening of pharmaceuticals using a genome-wide bacterial bioreporter array. Biosens Bioelectron. 2015;68:699–704.

  23. 23.

    Michelini E, Cevenini L, Calabretta MM, Calabria D, Roda A. Exploiting in vitro and in vivo bioluminescence for the implementation of the three Rs principle (replacement, reduction, and refinement) in drug discovery. Anal Bioanal Chem. 2014;406:5531–9.

  24. 24.

    Class B, Thorne N, Aguisanda F, Southall N, McKew JC, Zheng W. High-throughput viability assay using an autonomously bioluminescent cell line with a bacterial Lux reporter. J Lab Autom. 2015;20:164–74.

  25. 25.

    Raut N, O’Connor G, Pasini P, Daunert S. Engineered cells as biosensing systems in biomedical analysis. Anal Bioanal Chem. 2012;402:3147–59.

  26. 26.

    Michelini E, Cevenini L, Mezzanotte L, Leskinen P, Virta M, Karp M, et al. A sensitive recombinant cell-based bioluminescent assay for detection of androgen-like compounds. Nat Protoc. 2008;3:1895–902.

  27. 27.

    Branchini BR, Southworth TL, Fontaine DM, Kohrt D, Talukder M, Michelini E, et al. An enhanced chimeric firefly luciferase-inspired enzyme for ATP detection and bioluminescence reporter and imaging applications. Anal Biochem. 2015;484:148–53.

  28. 28.

    Branchini BR, Southworth TL, Fontaine DM, Davis AL, Behney CE, Murtiashaw MHA. Photinus pyralis and Luciola italica chimeric firefly luciferase produces enhanced bioluminescence. Biochemistry. 2014;53:6287–9.

  29. 29.

    Yasunaga M, Murotomi K, Abe H, Yamazaki T, Nishii S, Ohbayashi T, et al. Highly sensitive luciferase reporter assay using a potent destabilization sequence of calpain 3. J Biotechnol. 2015;194:115–23.

  30. 30.

    Ohmiya Y. Simultaneous multicolor luciferase reporter assays for monitoring of multiple genes expressions. Comb Chem High Throughput Screen. 2015;18:937–45.

  31. 31.

    Hall MP, Unch J, Binkowski BF, Valley MP, Butler BL, Wood MG, et al. Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol. 2012;7:1848–57.

  32. 32.

    Masser AE, Kandasamy G, Kaimal JM, Andréasson C. Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae. Yeast. 2016;33:191–200.

  33. 33.

    De Niz M, Stanway RR, Wacker R, Keller D, Heussler VT. An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle. Malar J. 2016;15:232.

  34. 34.

    Dixon AS, Schwinn MK, Hall MP, Zimmerman K, Otto P, Lubben TH, et al. NanoLuc complementation reporter optimized for accurate measurement of protein interactions in cells. ACS Chem Biol. 2016;11:400–8.

  35. 35.

    Boute N, Lowe P, Berger S, Malissard M, Robert A, Tesar M. NanoLuc luciferase—a multifunctional tool for high throughput antibody screening. Front Pharmacol. 2016;7:27.

  36. 36.

    England CG, Ehlerding EB, Cai W. NanoLuc: a small luciferase is brightening up the field of bioluminescence. Bioconjug Chem. 2016;27:1175–87.

  37. 37.

    Arts R, den Hartog I, Zijlema SE, Thijssen V, van der Beelen SH, Merkx M. Detection of antibodies in blood plasma using bioluminescent sensor proteins and a smartphone. Anal Chem. 2016;88:4525–32.

  38. 38.

    Fahrmann JF, Ballester OF, Ballester G, Witte TR, Salazar AJ, Kordusky B, et al. Inhibition of nuclear factor kappa B activation in early-stage chronic lymphocytic leukemia by omega-3 fatty acids. Cancer Invest. 2013;31:24–38.

  39. 39.

    Bak YK, Lampe JW, Sung MK. Effects of dietary supplementation of glucosamine sulfate on intestinal inflammation in a mouse model of experimental colitis. J Gastroenterol Hepatol. 2014;29:957–63.

  40. 40.

    Sánchez-Fidalgo S, Villegas I, Rosillo MÁ, Aparicio-Soto M, de la Lastra CA. Dietary squalene supplementation improves DSS-induced acute colitis by downregulating p38 MAPK and NFkB signaling pathways. Mol Nutr Food Res. 2015;59:284–92.

  41. 41.

    Ferri M, Bin S, Vallini V, Fava F, Michelini E, Roda A, et al. Recovery of polyphenols from red grape pomace and assessment of their antioxidant and anti-cholesterol activities. N Biotechnol. 2016;33:338–44.

  42. 42.

    Cevenini L, Calabretta MM, Calabria D, Roda A, Michelini E. Luciferase genes as reporter reactions: how to use them in molecular biology? Adv Biochem Eng Biotechnol. 2016;154:3–17.

  43. 43.

    Jara-Palacios MJ, Hernanz D, Cifuentes-Gomez T, Escudero-Gilete ML, Heredia FJ, Spencer JP. Assessment of white grape pomace from winemaking as source of bioactive compounds, and its antiproliferative activity. Food Chem. 2015;183:78–82.

  44. 44.

    Nunes C, Ferreira E, Freitas V, Almeida L, Barbosa RM, Laranjinha. Intestinal anti-inflammatory activity of red wine extract: unveiling the mechanisms in colonic epithelial cells. J.Food Funct. 2013;4:373-383

Download references


The grape pomace samples were obtained in the frame of a project supported by the Sadam Engineering (Sadam Eridania Spa group, Bologna, Italy) to Annalisa Tassoni. This research was sponsored in part by the NATO Science for Peace and Security Programme under Grant No. 985042.

Author information

Correspondence to Aldo Roda or Elisa Michelini.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Published in the topical collection Highlights of Analytical Chemical Luminescence with guest editors Aldo Roda, Hua Cui, and Chao Lu.

Electronic supplementary material

Below is the link to the electronic supplementary material.


(PDF 853 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Cevenini, L., Calabretta, M.M., Lopreside, A. et al. Exploiting NanoLuc luciferase for smartphone-based bioluminescence cell biosensor for (anti)-inflammatory activity and toxicity. Anal Bioanal Chem 408, 8859–8868 (2016). https://doi.org/10.1007/s00216-016-0062-3

Download citation


  • Bioluminescence
  • Smartphone
  • Anti-inflammatory activity
  • Toxicity
  • Portable analytical device
  • Cell biosensor