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Combined Poly(Lactide-Co-Glycolide) Microspheres Containing Diphtheria Toxoid for a Single-shot Immunization

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Abstract

To develop a single-shot vaccine containing diphtheria toxoid (DT) with a sufficient immune response, poly(lactide-co-glycolide) (PLGA) microspheres were prepared by water-in-oil-in-water double emulsification and solvent extraction techniques using low or high-molecular-weight PLGA (LMW-MS or HMW-MS). Stearic acid (SA) was introduced to HMW-MS (HMW/SA-MS) as a release modulator. Mean particle sizes (dvs, μm) varied between the prepared microspheres, with LMW-MS, HMW-MS, and HMW/SA-MS having the sizes of 29.83, 110.59, and 69.5 μm, respectively; however, the protein entrapment and loading efficiency did not vary, with values of 15.2–16.8 μg/mg and 61–75%, respectively. LMW-MS showed slower initial release (~ 2 weeks) but faster and higher release of antigen during weeks 3~7 than did HMW-MS. HMW/SA-MS showed rapid initial release followed by a continuous release over an extended period of time (~ 12 weeks). Mixed PLGA microspheres (MIX-MS), a combination of HMW/SA-MS and LMW-MS (1:1), demonstrated a sufficient initial antigen release and a subsequent boost release in a pulsatile manner. Serum antibody levels were measured by ELISA after DT immunization of Balb/c mice, and showed a greater response to MIX-MS than to alum-adsorbed DT (control). A lethal toxin challenge test with MIX-MS (a DT dose of 18 Lf) using Balb/c mice revealed complete protection, indicating a good candidate delivery system for a single-shot immunization.

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Funding

This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HI17C0710).

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Author notes

  1. Hye Seung Woo and Sung Rae Kim contributed equally to this work and should be considered as co-first authors.

Authors and Affiliations

  1. College of Pharmacy, Chung-Ang University, 84 Heuksuk-ro, Dongjak-gu, Seoul, 06974, South Korea

    Hye Seung Woo, Sung Rae Kim, Mikyeong Yoon, Eun Seok Lee, Do Ik Lee & Young Wook Choi

  2. Colleg of Medicine, Chung-Ang University, 84 Heuksuk-ro, Dongjak-gu, Seoul, 06974, South Korea

    In Ho Chang & Young Mi Whang

  3. College of Pharmacy, Dankook University, 119 Dandae-ro, Dongnam-gu, Cheonan, Chungnam, 31116, South Korea

    Myung Joo Kang

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  1. Hye Seung Woo
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Correspondence to Young Wook Choi.

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Woo, H.S., Kim, S.R., Yoon, M. et al. Combined Poly(Lactide-Co-Glycolide) Microspheres Containing Diphtheria Toxoid for a Single-shot Immunization. AAPS PharmSciTech 19, 1160–1167 (2018). https://doi.org/10.1208/s12249-017-0934-7

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  • DOI: https://doi.org/10.1208/s12249-017-0934-7

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