• Diky Mudhakir Department of , School of Pharmacy, Institut Teknologi Bandung, Bandung, Indonesia.
  • Adik Ahmadi Department of , School of Pharmacy, Institut Teknologi Bandung, Bandung, Indonesia.
  • Muhamad Insanu Department of , School of Pharmacy, Institut Teknologi Bandung, Bandung, Indonesia.
  • Neny Nuraini Biofarma Limited Company, Bandung, Indonesia.


Objective: Oftentimes, the recombinant antigen for the use of vaccines is less immunogenic than live attenuated or inactive vaccines. Hence, a potent adjuvant is needed to enhance the immune response. Moreover, the role of vector design is also important to facilitate the improvement of the immune response. The aim of this research was to develop hepatitis B surface antigen (HBsAg)-loaded nanoparticles and Moringa oleifera aqueous leaf extracts as an adjuvant using chitosan polymer.

Methods: Chitosan nanoparticles were prepared by the ionic gelation method using sodium tripolyphosphate as the cross-linking agent. A system was composed of chitosan core in which HBsAg and M. oleifera extracts were incorporated. The concentration of HBsAg used in this combination was 10 μg/ml, and the concentrations of extracts were 10, 50, and 100 μg/ml, respectively. In this study, three types of nanoparticles were produced: HBsAg-loaded nanoparticles, M. oleifera-loaded nanoparticles, and combination of HBsAg–M. oleifera-loaded nanoparticles. The nanoparticles formed were characterized by the particle size, HBsAg entrapment efficiency using sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the entrapment efficiency of extracts using the total flavonoid method.

Results: The results showed that the particle size was between 111 and 245 nm. The entrapment efficiency of HBsAg in the separate formula was 79%, while that in the combined formula was approximately 96–98%. Furthermore, the entrapment efficiency of the extracts in the separate formula was around 64–91%, while that in the combined formula was 55–82.5%. Particularly, HBsAg–M. oleifera-loaded chitosan nanoparticles with the extract concentrations of 50 μg/ml showed the highest entrapment efficiencies of HBsAg and M. oleifera extracts of approximately 98 and 82.5%, respectively.

Conclusion: Collectively, the system has been successfully developed, so it is then plausible to determine the function of the devices to enhance the immune response in the future.

Keywords: Nanoparticle, Chitosan, Hepatitis B surface antigen, Adjuvant, Moringa oleifera.


1. Arora D, Khurana B, Kumar MS, Vyas S. Oral immunization againts hepatitis B virus using mannosylated bilosomes. Int J Rec Adv Pharm Res 2011;1:45-51.
2. Zho F, Neutra MR. Antigen delivery to mucosa-associated lymphoid tissues using liposomes as a carrier. Biosci Rep 2002;22:355-69.
3. Das S, Desmukh R. Advances in vaccination: A review. Int J Appl Pharm 2009;1:1-21.
4. Galuh AS. Pengaruh pemberian ekstrak Phyllanthus niruri L terhadap fungsifagositosismakrofag pada mencitBALb/c yang diinfeksi Salmonella typhimurium. Semarang: Fakultas Kedokteran Universitas Diponegoro; 2008.
5. Rachmawati I, Muhaimin R. In vitro immunomodulatory activity of aqueous extract of Moringa oleifera Lam. Leaf to CD4+, CD8+, B220+ cells in Mus musculus. J Exp Life Sci 2014;4:15-20.
6. Hani N, Azarian MH, Torkamani AH, Mahmood WA. Characterization of gelatin nanoparticles encapsulated with Moringa oleifera bioactive extract. Int J Food Sci Technol 2016;51:2327-37.
7. Moodley JS, Krishna SB, Pillay K, Sershen, Govender P. Green synthesis of silver nanoparticles from Moringa oleifera leaf extracts and its antimicrobial potential. Adv Nat Sci Nanosci Nanotechnol 2018;9:1-9.
8. Choi Y, Kim HA, Kim KW, Lee BT. Comparative toxicity of silver nanoparticles and silver ions to Escherichia coli. J Environ Sci 2018;66:50-60.
9. Vongsak B, Sithisarn S, Mangmool S, Thongpraditchote S, Wongkrajang Y, Gritsanapan W. Maximizing total phenolics, total flavonoid contents, and antioxidant activity of Moringa oleifera leaf extract by the appropriate extract method. Ind Corps Prod 2012;44:566 71.
10. Bennet RN, Mellon FA, Foidl N, Pratt JH, Dupont MS, Perkins L, et al. Profiling glucosinolates and phenolics in vegetative and reproductive tissues of multi-purpose trees Moringa oleifera Lam. and Moringa stenopetala L. J Agric Food Chem 2003;51:3546-53.
11. Bose CK. Possible role of Moringa oleifera Lam root in epithelial ovarian cancer. Med Gen Med 2004;9:26.
12. Anwar F, Latiff S, Ashraf M, Gillani AH. Moringa oleifera: A food plant with multiple medicinal uses. Phytother Res 2007;21:17-25.
13. Sarvesh K, Kumar MS, Ankit S, Kumar SA. Quality control standardization of the bark of Moringa oleifera Lam. Int J Pharm Pharm Sci 2015;7:34-7.
14. Tiyaboonchai W. Chitosan nanoparticles: A promising system for drug delivery. Naresuan Univ J 2003;11:51-66.
15. Diminsky D, Schirmbeck R, Reimann J, Barenholz Y. Comparison between HBsAg particles derived from mammalian cells (CHO) and yeast cells: Composition, structure and immunogenicity. Vaccine 1997;15:637-47.
16. Hu B, Pan C, Sun Y, Hou Z, Ye H, Zeng X. Optimization of fabrication parameters to produce chitosan-tripolyphosphate nanoparticles for delivery of tea catechin. J Agric Food Chem 2008;56:7451-8.
17. Ruby JJ, Pandey VP. Chitosan nanoparticles as a nasal drug delivery for memantine hydrochloride. Int J Pharm Pharm Sci 2015;7:56-60.
18. Fan W, Yan W, Xu Z, Ni H. Formation mechanism of monodisperse, low molecular weight chitosan nanoparticles by ionic gelation technique. Colloid Surf B 2012;90:21-7.
19. Jonassen H, Kjeniksen AL, Hiorth M. Effects of ionic strength on the size and compactness of chitosan nanoparticles. Colloid Polym Sci 2012;290:919-29.
20. Kaloti M, Bohidar HB. Kinetics of coacervation transition versus nanoparticle formation in chitosan-sodium tripolyphosphate solutions. Colloid Surf B 2010;81:165-75.
21. Slütter B, Plapied L, Fievez V, Sande MA, des Rieux A, Schneider YJ, et al. Mechanistic study of the adjuvant effect of biodegradable nanoparticles in mucosal vaccination. J Control Release 2009;138:113 21.
22. Gao L, Zhang D, Chen M. Drug nanocrystals for formulation of poorly soluble drugs and its application as potential drug delivery system. J Nanopart Res 2008;10:845-62.
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How to Cite
Mudhakir, D., A. Ahmadi, M. Insanu, and N. Nuraini. “PACKAGING OF HEPATITIS B SURFACE ANTIGEN VACCINE AND MORINGA OLEIFERA EXTRACT INTO CHITOSAN NANOPARTICLES”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 1, Jan. 2019, pp. 346-50, doi:10.22159/ajpcr.2019.v12i1.15386.
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