Skip to main content
SpringerLink
Log in

A Simple Paper-Based α-Amylase Separating System for Potential Application in Biological Sciences

  • Original Article
  • Published:
BioChip Journal Aims and scope Submit manuscript

Abstract

The salivary glands are closely associated with stress conditions and the neuroendocrine system. The concentrations of these salivary biomarkers fluctuate with a person’s disease status. Until now, salivary diagnostics have been promising; however, the analytical tools for detecting salivary biomarkers have remained burdensome due to the lack of user-friendly and cost-effective equipment. In this study, we used a microfluidic starch-coated paper-based chip to isolate ɑ-amylase from human saliva. The concentration of ɑ-amylase on the head and tail sections of the paper-based chip was compared in the presence or absence of starch. We found that ɑ-amylase was highly concentrated only in the head section of the starch-coated paper-based chip through Western blotting (WB) and enzyme-linked immunosorbent assay (ELISA). We expect our designed paper-based chip will aid in α-amylase detection technologies using saliva and small amounts of other biofluids including blood, tears, pancreatic juice, and urine.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

References

  1. Bozovic, D., Racic, M., Ivkovic, N.: Salivary cortisol levels as a biological marker of stress reaction. Med. Arch. 67(5), 374–377 (2013)

    Article  Google Scholar 

  2. Beiter, R., Nash, R., McCrady, M., Rhoades, D., Linscomb, M., Clarahan, M., Sammut, S.: The prevalence and correlates of depression, anxiety, and stress in a sample of college students. J. Affective Disord. 173, 90–96 (2014)

    Article  Google Scholar 

  3. Vineetha, R., Pai, K.-M., Vengal, M., Gopalakrishna, K., Narayanakurup, D.: Usefulness of salivary alpha amylase as a biomarker of chronic stress and stress related oral mucosal changes—a pilot study. J. Clin. Exp. Dent. 6(2), e132–e137 (2014)

    Article  Google Scholar 

  4. Lima, D.P., Diniz, D.G., Moimaz, S.A.S., Sumida, D.H., Okamoto, A.C.: Saliva: reflection of the body. Int. J. Infect. Dis. 14(3), E184–E188 (2010)

    Article  Google Scholar 

  5. Granger, D.A., Kivlighan, K.T., El-Sheikh, M., Gordis, E.B., Stroud, L.R.: Salivary α-amylase in biobehavioral research. Ann. N. Y. Acad. Sci. 1098(1), 122–144 (2007)

    Article  CAS  Google Scholar 

  6. Capranica, L., Condello, G., Tornello, F., Iona, T., Chiodo, S., Valenzano, A., De Rosas, M., Messina, G., Tessitore, A., Cibelli, G.: Salivary alpha-amylase, salivary cortisol, and anxiety during a youth taekwondo championship: an observational study. Medicine (Baltimore) 96(28), e7272–e7272 (2017)

    Article  CAS  Google Scholar 

  7. Nater, U.M., Rohleder, N., Gaab, J., Berger, S., Jud, A., Kirschbaum, C., Ehlert, U.: Human salivary alpha-amylase reactivity in a psychosocial stress paradigm. Int. J. Psychophysiol. 55(3), 333–342 (2005)

    Article  Google Scholar 

  8. Wolf, J.M., Nicholls, E., Chen, E.: Chronic stress, salivary cortisol, and α-amylase in children with asthma and healthy children. Biol. Psychol. 78(1), 20–28 (2008)

    Article  Google Scholar 

  9. Tanaka, Y., Maruyama, Y., Ishitobi, Y., Kawano, A., Ando, T., Ikeda, R., Inoue, A., Imanaga, J., Okamoto, S., Kanehisa, M., Ninomiya, T., Tsuru, J., Akiyoshi, J.: Salivary alpha-amylase and cortisol responsiveness following electrically stimulated physical stress in bipolar disorder patients. Neuropsychiatr. Dis. Treat. 9, 1899–1905 (2013)

    PubMed  PubMed Central  Google Scholar 

  10. Cozma, S., Dima-Cozma, L.C., Ghiciuc, C.M., Pasquali, V., Saponaro, A., Patacchioli, F.R.: Salivary cortisol and α-amylase: subclinical indicators of stress as cardiometabolic risk. Braz. J. Med. Biol. Res. 50(2), e5577–e5577 (2017)

    Article  CAS  Google Scholar 

  11. Liu, X.Y., Cheng, C.M., Martinez, A.W., Mirica, K.A., Li, X.J., Phillips, S.T., Mascareñas, M., Whitesides, G.M.: In A portable microfluidic paper-based device for ELISA, 2011 IEEE 24th International Conference on Micro Electro Mechanical Systems, pp 75–78 (2011)

  12. Nilghaz, A., Guan, L., Tan, W., Shen, W.: Advances of paper-based microfluidics for diagnostics—the original motivation and current status. ACS Sens. 1(12), 1382–1393 (2016)

    Article  CAS  Google Scholar 

  13. He, Y., Wu, Y., Fu, J.-Z., Wu, W.-B.: Fabrication of paper-based microfluidic analysis devices: a review. RSC Adv. 5(95), 78109–78127 (2015)

    Article  CAS  Google Scholar 

  14. Sharma, N., Barstis, T., Giri, B.: Advances in paper-analytical methods for pharmaceutical analysis. Eur. J. Pharm. Sci. 111, 46–56 (2018)

    Article  CAS  Google Scholar 

  15. Oyola-Reynoso, S., Heim, A.P., Halbertsma-Black, J., Zhao, C., Tevis, I.D., Çınar, S., Cademartiri, R., Liu, X., Bloch, J.-F., Thuo, M.M.: Draw your assay: fabrication of low-cost paper-based diagnostic and multi-well test zones by drawing on a paper. Talanta 144, 289–293 (2015)

    Article  CAS  Google Scholar 

  16. Lee, J.-H., Jung, H.-I.: Biochip technology for monitoring posttraumatic stress disorder (PTSD). BioChip J. 7(3), 195–200 (2013)

    Article  CAS  Google Scholar 

  17. Pandey, C., Augustine, S., Kumar, S., Kumar, S., Nara, S., Srivastava, S., Malhotra, B.: Microfluidics based point‐of‐care diagnostics. Biotechnol. J. 13 (2017)

  18. Songjaroen, T., Dungchai, W., Chailapakul, O., Laiwattanapaisal, W.: Novel, simple and low-cost alternative method for fabrication of paper-based microfluidics by wax dipping. Talanta 85(5), 2587–2593 (2011)

    Article  CAS  Google Scholar 

  19. Elizalde, E., Urteaga, R., Berli, C.: Precise capillary flow for paper-based viscometry. Microfluidics Nanofluidics 20 (2016)

  20. Chun, H.J., Park, Y.M., Han, Y.D., Jang, Y.H., Yoon, H.C.: Paper-based glucose biosensing system utilizing a smartphone as a signal reader. BioChip J. 8(3), 218–226 (2014)

    Article  CAS  Google Scholar 

  21. Choi, R.J., Yong, W.K., Choi, Y.J., Cowie, C.A.: Emerging point-of-care technologies for food safety analysis. Sensors. 19 (4) (2019)

  22. Dungchai, W., Chailapakul, O., Henry, C.S.: Electrochemical detection for paper-based microfluidics. Anal. Chem. 81(14), 5821–5826 (2009)

    Article  CAS  Google Scholar 

  23. Gong, M.M., Sinton, D.: Turning the page: advancing paper-based microfluidics for broad diagnostic application. Chem. Rev. 117(12), 8447–8480 (2017)

    Article  CAS  Google Scholar 

  24. Martinez, A.W., Phillips, S.T., Whitesides, G.M., Carrilho, E.: Diagnostics for the developing world: microfluidic paper-based analytical devices. Anal. Chem. 82(1), 3–10 (2010)

    Article  CAS  Google Scholar 

  25. Deutsch, O., Fleissig, Y., Zaks, B., Krief, G., Aframian, D.J., Palmon, A.: An approach to remove alpha amylase for proteomic analysis of low abundance biomarkers in human saliva. Electrophoresis 29(20), 4150–4157 (2008)

    Article  CAS  Google Scholar 

  26. Nater, U.M., La Marca, R., Erni, K., Ehlert, U.: Alpha-amylase activity in blood increases after pharmacological, but not psychological, activation of the adrenergic system. PLoS ONE 10(6), e0130449 (2015)

    Article  Google Scholar 

  27. van Haeringen, N.J., Ensink, F., Glasius, E.: Amylase in human tear fluid: origin and characteristics, compared with salivary and urinary amylases. Exp. Eye Res. 21(4), 395–403 (1975)

    Article  Google Scholar 

  28. Shin, S., Kim, N., Hong, J.W.: Comparison of surface modification techniques on polydimethylsiloxane to prevent protein adsorption. BioChip J. 12(2), 123–127 (2018)

    Article  CAS  Google Scholar 

  29. Khurshid, Z., Zohaib, S., Najeeb, S., Zafar, M.S., Slowey, P.D., Almas, K.: Human saliva collection devices for proteomics: an update. Int J Mol Sci 17(6), 846 (2016)

    Article  Google Scholar 

  30. Zhang, W., Du, Y., Wang, M.L.: Noninvasive glucose monitoring using saliva nano-biosensor. Sens. Bio-Sens. Res. 4, 23–29 (2015)

    Article  Google Scholar 

  31. Lee, M., Ban, J.-J., Im, W., Kim, M.: Influence of storage condition on exosome recovery. Biotechnol. Bioproc. Eng. 21(2), 299–304 (2016)

    Article  CAS  Google Scholar 

  32. Herr, A.E., Hatch, A.V., Throckmorton, D.J., Tran, H.M., Brennan, J.S., Giannobile, W.V., Singh, A.K.: Microfluidic immunoassays as rapid saliva-based clinical diagnostics. Proc. Natl. Acad. Sci. 104(13), 5268 (2007)

    Article  CAS  Google Scholar 

  33. Khan, R.S., Khurshid, Z., Yahya Ibrahim Asiri, F.: Advancing point-of-care (PoC) testing using human saliva as liquid biopsy. Diag. Basel 7(3), 39 (2017)

    Google Scholar 

  34. Vitorino, R., Lobo, M.J.C., Ferrer-Correira, A.J., Dubin, J.R., Tomer, K.B., Domingues, P.M., Amado, F.M.L.: Identification of human whole saliva protein components using proteomics. Proteomics 4(4), 1109–1115 (2004)

    Article  CAS  Google Scholar 

  35. Sultan, Z.: Salivary point-of-care technology. Eur. J. Dent. 12, 1 (2018)

    Article  Google Scholar 

  36. Noiphung, J., Nguyen, M.P., Punyadeera, C., Wan, Y., Laiwattanapaisal, W., Henry, C.S.: Development of paper-based analytical devices for minimizing the viscosity effect in human saliva. Theranostics 8(14), 3797–3807 (2018)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research was supported by Basic Science Research Program (2018R1A2B6005354) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, Future Planning of Korea, and National Research Foundation of Korea (NRF-2018M3A9H1023323).

Author information

Author notes

  1. Sujin Hyung and Gul Karima contributed equally to this study.

Authors and Affiliations

  1. Center for Exosome and Bioparticulate Research, Hanyang University, Ansan, Gyeonggi-do, 15588, Korea

    Sujin Hyung, Gul Karima, Kyusoon Shin, Kwang Sik Kim & Jong Wook Hong

  2. Department of Bionanotechnology, Graduate School, Hanyang University, Seoul, 04763, Korea

    Gul Karima, Kyusoon Shin, Kwang Sik Kim & Jong Wook Hong

  3. Department of Bionanoengineering, Hanyang University, Ansan, Gyeonggi-do, 15588, Korea

    Jong Wook Hong

Authors
  1. Sujin Hyung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gul Karima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kyusoon Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kwang Sik Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jong Wook Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jong Wook Hong.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 174 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hyung, S., Karima, G., Shin, K. et al. A Simple Paper-Based α-Amylase Separating System for Potential Application in Biological Sciences. BioChip J 15, 252–259 (2021). https://doi.org/10.1007/s13206-021-00022-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13206-021-00022-3

Keywords

Search

Navigation