Preparation and characterization of PLGA nanospheres loaded with inactivated influenza virus, CpG-ODN and Quillaja saponin


1 Student Research Committee, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Food and Drug Control, Students Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran

3 Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran



The purpose of this study was preparation and evaluation of PLGA nanospheres containing the influenza virus and different adjuvants, Quillaja saponin (QS) and CpG-ODN.
Materials and Methods:
Nanospheres were prepared using the double emulsion-solvent evaporation method. The morphological and physicochemical properties were studied by scanning electron microscopy (SEM), determination of zeta potential, encapsulation efficiency and release profile.
The particle size of formulations was less than 1000 nm, except for formulations containing antigen. The results were confirmed with SEM images. Encapsulation efficiency of antigen, QS and CpG ODN were 80%, 62% and 31%, respectively. The zeta potential of nanospheres was about -30 mV. The burst release was observed for all encapsulates and reached to about 48%, 44% and 35% within 90 min for antigen, CpG-ODN and Qs content, respectively.
The formulations showed proper physicochemical properties. These nanospheres have good potential to be used as delivery systems/adjuvants for immunization against influenza.


1. Carter J, Saunders V. Virology: Principles and Applications: John Wiley & Sons, Chichester; 2007.
2. Amorij JP, Huckriede A, Wilschut J, Frijlink HW, Hinrichs WLJ. Development of stable influenza vaccine powder formulations: Challenges and possibilities. Pharm Res 2008; 25:1256-1273.
3. Smith DM, Simon JK, Baker JR Jr. Applications of nanotechnology for immunology. Nat Rev Immunol 2013; 13:592-605.
4. Tafaghodi M, Eskandari M, Kharazizadeh M, Khamesipour A, Jaafari MR. Immunization against leishmaniasis by PLGA nanospheres loaded with an experimental autoclaved Leishmania major (ALM) and Quillaja saponins. Trop Biomed 2010; 27:639-650.
5. Mohaghegh M, Tafaghodi M. Dextran microspheres could enhance immune responses against PLGA nanospheres encapsulated with tetanus toxoid and Quillaja saponins after nasal immunization in rabbit. Pharm Dev Technol 2011; 16:36-43.
6. Babapoor S, Neef T, Mittelholzer C, Girshick T, Garmendia A, Shang H,
et al. A novel vaccine using nanoparticle platform to present immunogenic M2e against avian influenza infection. Influenza Res Treat 2011; 2011:126794.
7. Keijzer C, Slütter B, van der Zee R, Jiskoot W, van Eden W, Broere F. PLGA, PLGA-TMC and TMC-TPP nanoparticles differentially modulate the outcome of nasal vaccination by inducing tolerance or enhancing humoral immunity. PLoS One 2011; 6:e26684.
8. Zaman M, Chandrudu S, Toth I. Strategies for intranasal delivery of vaccines. Drug Deliv Transl Res 2013; 3:100-109.
9. Dehghan S, Tavassoti Kheiri M, Tabatabaiean M, Darzi S, Tafaghodi M. Dry-powder form of chitosan nanospheres containing influenza virus and adjuvants for nasal immunization. Arch Pharm Res 2013; 36:981-992.
Mohajer et al PLGA nanospheres promising nasal delivery systems Iran J Basic Med Sci, Vol. 17, No. 9, Sep 2014 5
10. Sahoo SK, Panyam J, Prabha S, Labhasetwar V. Residual polyvinyl alcohol associated with poly (D,L-lactide-co-glycolide) nanoparticles affects their physical properties and cellular uptake. J Control Release 2002; 82:105-114.
11. Alpar HO, Almeida AJ. Identification of some of the physico-chemical characteristics of microspheres which influence the induction of the immune response following mucosal delivery. Eur J Pharm Biopharm 1994; 40:198-202.
12. Khameneh B, Momen-Nejad M, Tafaghodi M.
In vivo evaluation of mucoadhesive properties of nanoliposomal formulations upon coating with trimethylchitosan polymer. Nanomed J 2014; 1:147-154.
13. Yang L, Chen L, Zeng R, Li C, Qiao R, Hu L,
et al. Synthesis, nanosizing and in vitro drug release of a novel anti-HIV polymeric prodrug: Chitosan-O-isopropyl-5′-O-d4T monophosphate conjugate. Bioorg Med Chem 2010; 18:117-123.
14. Xue SW. Synthesis, characterization, biodegradation, and drug delivery application of biodegradable lactic/glycolic acid polymers: Part III. Drug delivery application. Artif Cells Blood Substit Immobil Biotechnol 2004; 32:575-591.
15. Benelli P, Conti B, Genta I, Costantini M, Montanari L. Clonazepam microencapsulation in poly-D,L-lactide-co-glycolide microspheres. J Microencapsul 1998; 15:431-443.