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Modeling, simulation, and employing dilution–dialysis microfluidic chip (DDMC) for heightening proteins refolding efficiency

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Abstract

Miniaturized systems based on the principles of microfluidics are widely used in various fields, such as biochemical and biomedical applications. Systematic design processes are demanded the proper use of these microfluidic devices based on mathematical simulations. Aggregated proteins (e.g., inclusion bodies) in solution with chaotropic agents (such as urea) at high concentration in combination with reducing agents are denatured. Refolding methods to achieve the native proteins from inclusion bodies of recombinant protein relying on denaturant dilution or dialysis approaches for suppressing protein aggregation is very important in the industrial field. In this paper, a modeling approach is introduced and employed that enables a compact and cost-effective method for on-chip refolding process. The innovative aspect of the presented refolding method is incorporation dialysis and dilution. Dilution–dialysis microfluidic chip (DDMC) increases productivity folding of proteins with the gradual reduction of the amount of urea. It has shown the potential of DDMC for performing refolding of protein trials. The principles of the microfluidic device detailed in this paper are to produce protein on the dilution with slow mixing through diffusion of a denatured protein solution and stepwise dialysis of a refolding buffer flowing together and the flow regime is creeping flow. The operation of DDMC was modeled in two dimensions. This system simulated by COMSOL Multiphysics Modeling Software. The simulation results for a microfluidic refolding chip showed that DDMC was deemed to be perfectly suitable for control decreasing urea in the fluid model. The DDMC was validated through an experimental study. According to the results, refolding efficiency of denaturant Hen egg white lysozyme (HEWL) (EC 3.2.1.17) used as a model protein was improved. Regard to the remaining activity test, it was increased from 42.6 in simple dilution to 93.7 using DDMC.

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References

  1. Kashanian F, Kokkinis G, Bernardi J, Zand MR, Shamloo A, Giouroudi I (2018) A novel magnetic microfluidic platform for on-chip separation of 3 types of silica coated magnetic nanoparticles (Fe3O4@ SiO2). Sens Actuators A Phys 270:223–230

    Article  CAS  Google Scholar 

  2. Chen J, Chen D, Xie Y, Yuan T, Chen X (2013) Progress of microfluidics for biology and medicine. Nano-Micro Lett 5(1):66–80

    Article  CAS  Google Scholar 

  3. Yun KS, Lee D, Kim HS, Yoon E (2006) A microfluidic chip for measurement of biomolecules using a microbead-based quantum dot fluorescence assay. Meas Sci Technol 17(12):3178

    Article  CAS  Google Scholar 

  4. Yeo LY, Chang HC, Chan PP, Friend JR (2011) Microfluidic devices for bioapplications. Small 7(1):12–48

    Article  CAS  Google Scholar 

  5. Jungbauer A, Kaar W Current status of technical protein refolding. J Biotechnol 128(3):587–596

  6. Kashanian F, Habibi-Rezaei M, Bagherpour AR, Seyedarabi A, Moosavi-Movahedi AA (2017) Magnetic nanoparticles as double-edged swords: concentration-dependent ordering or disordering effects on lysozyme. RSC Adv 7(86):54813–54822

    Article  CAS  Google Scholar 

  7. Wang W, Nema S, Teagarden D (2010) Protein aggregation—pathways and influencing factors. Int J Pharm 390(2):89–99

    Article  CAS  Google Scholar 

  8. Singh SM, Panda AK Solubilization and refolding of bacterial inclusion body proteins. J Biosci Bioeng 99(4):303–310

  9. Singh A, Upadhyay V, Upadhyay AK, Singh SM, Panda AK (2015) Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process. Microb Cell Factories 14(1):41

    Article  Google Scholar 

  10. Clark EDB (2001) Protein refolding for industrial processes. Curr Opin Biotechnol 12(2):202–207

    Article  CAS  Google Scholar 

  11. Yamaguchi H, Miyazaki M, Briones-Nagata MP, Maeda H (2010) Refolding of difficult-to-fold proteins by a gradual decrease of denaturant using microfluidic chips. J Biochem 147(6):895–903

    Article  CAS  Google Scholar 

  12. Yamaguchi H, Miyazaki M (2015) Microfluidic chips with multi-junctions: an advanced tool in recovering proteins from inclusion bodies. Bioengineered 6(1):1–4

    Article  CAS  Google Scholar 

  13. Yamamoto E, Yamaguchi S, Sasaki N, Kim HB, Kitamori T, Nagamune T (2010) Artificial chaperone-assisted refolding in a microchannel. Bioprocess Biosyst Eng 33(1):171

    Article  CAS  Google Scholar 

  14. Zaccai NR, Yunus K, Matthews SM, Fisher AC, Falconer RJ (2007) Refolding of a membrane protein in a microfluidics reactor. Eur Biophys J 36(6):581–588

    Article  CAS  Google Scholar 

  15. Tsumoto K, Ejima D, Kumagai I, Arakawa T (2003) Practical considerations in refolding proteins from inclusion bodies. Protein Expr Purif 28(1):1–8

    Article  CAS  Google Scholar 

  16. Yamaguchi S, Yamamoto E, Mannen T, Nagamune T (2013) Protein refolding using chemical refolding additives. Biotechnol J 8(1):17–31

    Article  CAS  Google Scholar 

  17. Eiberle MK, Jungbauer A (2010) Technical refolding of proteins: Do we have freedom to operate?. Biotechnol J 5(6):547–559

    Article  CAS  Google Scholar 

  18. Umetsu M, Tsumoto K, Hara M, Ashish K, Goda S, Adschiri T, Kumagai I (2003) How additives influence the refolding of immunoglobulin-folded proteins in a stepwise dialysis system spectroscopic evidence for highly efficient refolding of a single-chain fv fragment. J Biol Chem 278(11):8979–8987

    Article  CAS  Google Scholar 

  19. Hashim U, Diyana PA, Adam T (2012) Numerical simulation of microfluidic devices. In: Semiconductor electronics (ICSE), 2012 10th IEEE international conference on 2012 Sep 19. IEEE, pp 26–29

  20. Chován T, Guttman A (2002) Microfabricated devices in biotechnology and biochemical processing. TRENDS Biotechnol 20(3):116–122

    Article  Google Scholar 

  21. Franke TA, Wixforth A (2008) Microfluidics for miniaturized laboratories on a chip. Chem Phys Chem 9(15):2140–2156

    Article  CAS  Google Scholar 

  22. Bian LJ, Dong FX, Liang CL, Yang XY, Liu L (2007) Studies on the refolding of egg white lysozyme denatured by urea using “phase diagram” method of fluorescence. Chin J Chem 25(12):1896–1903

    Article  CAS  Google Scholar 

  23. Liu Z, García-Díaz B, Catacchio B, Chiancone E, Vogel HJ (2015) Protecting Gram-negative bacterial cell envelopes from human lysozyme: interactions with Ivy inhibitor proteins from Escherichia coli and Pseudomonas aeruginosa. Biochim Biophys Acta (BBA) Biomembr 1848(11):3032–3046

    Article  CAS  Google Scholar 

  24. Feynman RP (1964) Feynman lectures on physics. In: Feynman RP, Leighton RB, Sands M (eds) Mainly electromagnetism and matter, vol 2. Addison-Wesley, Reading

    Google Scholar 

  25. Happel J, Brenner H (2012) Low Reynolds number hydrodynamics: with special applications to particulate media. Springer, New York

    Google Scholar 

  26. Atkinson B, Brocklebank MP, Card CC, Smith JM (1969) Low Reynolds number developing flows. AIChE J 15(4):548–553

    Article  CAS  Google Scholar 

  27. Aune KC, Tanford C (1969) Thermodynamics of the denaturation of lysozyme by guanidine hydrochloride. I. Dependence on pH at 25. Biochemistry 8(11):4579–4585

    Article  CAS  Google Scholar 

  28. Rozet E, Wascotte V, Lecouturier N, Préat V, Dewé W, Boulanger B, Hubert P (2007) Improvement of the decision efficiency of the accuracy profile by means of a desirability function for analytical methods validation: application to a diacetyl-monoxime colorimetric assay used for the determination of urea in transdermal iontophoretic extracts. Anal Chim Acta 591(2):239–247

    Article  CAS  Google Scholar 

  29. Shugar D (1952) The measurement of lysozyme activity and the ultra-violet inactivation of lysozyme. Biochim Biophys Acta 8:302–309

    Article  CAS  Google Scholar 

  30. Laurell T, Lenshof A (eds) (2014) Microscale acoustofluidics. Royal Society of Chemistry, p 8

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Authors and Affiliations

  1. Department of Life Science Engineering, Faculty of Disciplinary New Science and Technology, University of Tehran, Tehran, Iran

    F. Kashanian

  2. School of Biology, College of Science, University of Tehran, Tehran, Iran

    F. Kashanian & M. Habibi-Rezaei

  3. School of Mechanical Engineering, Sharif University of Technology, Tehran, Iran

    M. M. Masoudi & A. Shamloo

  4. Nano-Biomedicine Center of Excellence, Nanoscience and Nanotechnology Research Center, University of Tehran, Tehran, Iran

    M. Habibi-Rezaei

  5. Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran

    A. A. Moosavi-Movahedi

  6. Center of Excellence in Biothermodynamics, University of Tehran, Tehran, Iran

    A. A. Moosavi-Movahedi

Authors
  1. F. Kashanian
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  2. M. M. Masoudi
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  3. A. Shamloo
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  4. M. Habibi-Rezaei
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  5. A. A. Moosavi-Movahedi
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Correspondence to A. Shamloo or M. Habibi-Rezaei.

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Kashanian, F., Masoudi, M.M., Shamloo, A. et al. Modeling, simulation, and employing dilution–dialysis microfluidic chip (DDMC) for heightening proteins refolding efficiency. Bioprocess Biosyst Eng 41, 707–714 (2018). https://doi.org/10.1007/s00449-018-1904-5

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  • DOI: https://doi.org/10.1007/s00449-018-1904-5

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