The effect of SPION size and salting-out on transduction of PEGylated lentiviral vector

Document Type : Research Paper


1 Gene Therapy Research Center, Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran, Iran. Department of Materials Science and Engineering, Sharif University of Technology, Azadi Avenue, Tehran, Iran; SABZ Biomedicals Science-Based Company, Tehran, Iran;

2 Department of Materials Science and Engineering, Sharif University of Technology, Azadi Avenue, Tehran, Iran.

3 Ali-Asghar Children Hospital, Iran University of Medical Sciences, Tehran, Iran.

4 Department of Materials Science and Engineering, Sharif University of Technology, Azadi Avenue, Tehran, Iran. SABZ Biomedicals Science-Based Company, Tehran, Iran;

5 Gene Therapy Research Center, Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

6 Nanotechnology & Regenerative Medicine Commercialisation Centre (NanoRegMed Ltd, Nanoloom Ltd, & Liberum Health Ltd), London BioScience Innovation Centre, London, UK.


Gene editing has many promising applications for the treatment of diseases with unmet clinical needs, including cancers and autoimmune. There are two main routes for gene delivery: viral and non-viral. Recent research shows viral methods are emerging in clinical trials. Nevertheless, there are still a couple of technological obstacles that require further improvements, these include virus concentration and low efficiency of transduction. This research aimed to employ superparamagnetic iron oxide nanoparticles (SPION) to solve these problems. Three sizes of 10, 40, and 120 nm SPION were synthesized by co-precipitation and Sol-Gel methods, and characterized by XRD, FTIR, FESEM, and VSM. Conjugating SPION to viruses by polyethylene glycol (PEG) can increase the sedimentation of viruses due to magnetic and gravity forces even without ultracentrifuge. Moreover, this magnetic force can guide viruses toward cells and tremendously facilitate the transduction process. As shown, average size SPION (40 nm) revealed the best performance, especially in combination with salting-out precipitation and increased transduction efficiency of more than 20-fold. SPION size has significant effects and should be considered for this application. The combination of the salting-out method and SPION has a synergic effect and elevates transduction results tremendously.


  1. Kuwana Y, Asakura Y, Utsunomiya N, Nakanishi M, Arata Y, Itoh S, et al. Expression of chimeric receptor composed of immunoglobulin-derived V resions and T-cell receptor-derived C regions. Biochemical and Biophysical Research Communications. 1987;149(3):960-8.
  2. June CH, Ledbetter JA, Gillespie MM, Lindsten T, Thompson CB. T-Cell Proliferation Involving the CD28 Pathway Is Associated witH Cyclosporine-Resistant Interleukin 2 Gene Expression. Molecular and Cellular Biology. 1987;7(12):4472-81.
  3. Tebas P, Stein D, Tang WW, Frank I, Wang SQ, Lee G, et al. Gene editing of CCR5 in autologous CD4 T cells of persons infected with HIV. N Engl J Med. 2014;370(10):901-10.
  4. Sharma R, Anguela XM, Doyon Y, Wechsler T, DeKelver RC, Sproul S, et al. In vivo genome editing of the albumin locus as a platform for protein replacement therapy. Blood. 2015;126(15):1777-84.
  5. Luo D, Saltzman WM. Enhancement of transfection by physical concentration of DNA at the cell surface. Nature Biotechnology. 2000;18(8):893-5.
  6. Bishop NE, Anderson DA. Early interactions of hepatitis A virus with cultured cells: viral elution and the effect of pH and calcium ions. Archives of Virology. 1997;142(11):2161-78.
  7. Rivière GN, Korpi A, Sipponen MH, Zou T, Kostiainen MA, Österberg M. Agglomeration of Viruses by Cationic Lignin Particles for Facilitated Water Purification. ACS Sustain Chem Eng. 2020;8(10):4167-77.
  8. Saito K, Otsuki N, Takeda M, Hanada K. Liposome Flotation Assay for Studying Interactions Between Rubella Virus Particles and Lipid Membranes. Bio Protoc. 2018;8(16):e2983-e.
  9. Cearley CN, Wolfe JH. A single injection of an adeno-associated virus vector into nuclei with divergent connections results in widespread vector distribution in the brain and global correction of a neurogenetic disease. Journal of Neuroscience. 2007 Sep 12;27(37):9928-40.
  10. Thomas J-L, Bardou J, L'Hoste S, Mauchamp B, Chavancy G. A helium burst biolistic device adapted to penetrate fragile insect tissues. Journal of Insect Science. 2001;1(9):1-10.
  11. Kurita H, Takahashi S, Asada A, Matsuo M, Kishikawa K, Mizuno A, Numano R. Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor. PLoS One. 2015;10(12):e0144254-e.
  12. Lissandrello CA, Santos JA, Hsi P, Welch M, Mott VL, Kim ES, et al. High-throughput continuous-flow microfluidic electroporation of mRNA into primary human T cells for applications in cellular therapy manufacturing. Sci Rep. 2020;10(1):18045-.


  1. Mahmoudi M, Sant S, Wang B, Laurent S, Sen T. Superparamagnetic iron oxide nanoparticles (SPIONs): Development, surface modification and applications in chemotherapy. Advanced Drug Delivery Reviews. 2011;63(1-2):24-46.
  2. Allard-Vannier E, Hervé-Aubert K, Kaaki K, Blondy T, Shebanova A, Shaitan KV, et al. Folic acid-capped PEGylated magnetic nanoparticles enter cancer cells mostly via clathrin-dependent endocytosis. Biochimica et Biophysica Acta (BBA) - General Subjects. 2017;1861(6):1578-86.
  3. Ahmadi R, Hosseini HRM, Masoudi A, Omid H, Namivandi-Zangeneh R, Ahmadi M, et al. Effect of concentration on hydrodynamic size of magnetite-based ferrofluid as a potential MRI contrast agent. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 2013;424:113-7.
  4. Alromi DA, Madani SY, Seifalian A. Emerging Application of Magnetic Nanoparticles for Diagnosis and Treatment of Cancer. Polymers (Basel). 2021;13(23):4146.
  5. Wong J, Prout J, Seifalian A. Magnetic Nanoparticles: New Perspectives in Drug Delivery. Current Pharmaceutical Design. 2017;23(20).
  6. Scherer F, Anton M, Schillinger U, Henke J, Bergemann C, Krüger A, et al. Magnetofection: enhancing and targeting gene delivery by magnetic force in vitro and in vivo. Gene Therapy. 2002;9(2):102-9.
  7. Iwasaki T, Mizutani N, Watano S, Yanagida T, Kawai T. Size control of magnetite nanoparticles by organic solvent-free chemical coprecipitation at room temperature. Journal of Experimental Nanoscience. 2010;5(3):251-62.
  8. Sugimoto T, Matijević E. Formation of uniform spherical magnetite particles by crystallization from ferrous hydroxide gels. Journal of Colloid and Interface Science. 1980;74(1):227-43.
  9. Gheisari Y, Azadmanesh K, Ahmadbeigi N, Nassiri SM, Golestaneh AF, Naderi M, et al. Genetic Modification of Mesenchymal Stem Cells to Overexpress CXCR4 and CXCR7 Does Not Improve the Homing and Therapeutic Potentials of These Cells in Experimental Acute Kidney Injury. Stem Cells and Development. 2012;21(16):2969-80.
  10. Cullity BD. Elements of X-ray Diffraction. Addison-Wesley Publishing; 1956.
  11. Gupta AK, Wells S. Surface-Modified Superparamagnetic Nanoparticles for Drug Delivery: Preparation, Characterization, and Cytotoxicity Studies. IEEE Transactions on Nanobioscience. 2004;3(1):66-73.
  12. Hu L, Hach D, Chaumont D, Brachais CH, Couvercelle JP. One step grafting of monomethoxy poly(ethylene glycol) during synthesis of maghemite nanoparticles in aqueous medium. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 2008;330(1):1-7.
  13. Kim M, Jung J, Lee J, Na K, Park S, Hyun J. Amphiphilic comblike polymers enhance the colloidal stability of Fe3O4 nanoparticles. Colloids and Surfaces B: Biointerfaces. 2010;76(1):236-40.
  14. Lentivirus Gene Engineering Protocols. Methods in Molecular Biology: Humana Press; 2010.
  15. Yamada K, McCarty DM, Madden VJ, Walsh CE. Lentivirus Vector Purification Using Anion Exchange HPLC Leads to Improved Gene Transfer. BioTechniques. 2003;34(5):1074-80.
  16. Jiang W, Hua R, Wei M, Li C, Qiu Z, Yang X, Zhang C. An optimized method for high-titer lentivirus preparations without ultracentrifugation. Sci Rep. 2015;5:13875-.