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A cell ELISA based method for exosome detection in diagnostic and therapeutic applications

  • Shabanali Khodashenas
  • Saeed Khalili
  • Mehdi Forouzandeh MoghadamEmail author
Original Research Paper

Abstract

Objective

Exosomes are nano-scaled carriers of miRNA, mRNA and proteins which are secreted from viable cells. Exosome detection within serum, saliva and semen offers diagnostic value for detection of various diseases including cancer. In the present study, we have lunched an in vitro study to develop a more efficient method for exosome detection. In this regard, the recombinant LAMP-DARPins (capable of Her2 targeting) gene was packed within the lentiviral particles and stably transferred into the HEK293T cells. The morphology and sizes of the obtained exosomes were characterized by TEM and zeta sizer. The expression of LAMP-DARPins antigen on the exosome surface was verified by western blotting. Ultimately, the efficiency of cell surface ELISA in suspension method was examined for exosome detection.

Results

The exosome particles were successfully harvested and purified from transfected cells. The sizes of exosome particles were determined to be 90 nm using zeta sizer instrument. The TEM scan showed that the exosomes are cap like shaped and their sizes range between 40 and 150 nm. An observed 120 kDa band on western blotting paper indicated the LAMP-DARPins antigen expression on exosome surfaces. The results of cell surface ELISA in suspension method were superior to the results of conventional cell ELISA.

Conclusions

It could be concluded that the cell surface ELISA in suspension method could be an amenable method to detect exosome particles within the biological samples. Moreover, the method could be modified to evaluate the ability of exosomes to interact with target cells in both diagnostic and therapeutic applications.

Keywords

Cell surface ELISA in suspension DARPins Exosome LAMP2 

Notes

Acknowledgements

The authors wish to thank Tarbiat Modares University and Iranian National Science Foundation for supporting the conduct of this research.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Shabanali Khodashenas
    • 1
    • 2
    • 4
  • Saeed Khalili
    • 3
    • 4
  • Mehdi Forouzandeh Moghadam
    • 4
    Email author
  1. 1.Molecular and Cell Biology Research Center, Faculty of MedicineMazandaran University of Medical SciencesSariIran
  2. 2.Immunogenetics Research CenterMazandaran University of Medical SciencesSariIran
  3. 3.Department of Biology SciencesShahid Rajaee Teacher Training UniversityTehranIran
  4. 4.Department of Medical Biotechnology, Faculty of Medical SciencesTarbiat Modares UniversityTehranIran

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