標題: 運用奈米金粒子作為高效率的胜肽疫苗載體
Assessment for the size-dependent properties of gold nanoparticles as vaccine carrier to elicit focused and enhanced antibody response against synthetic peptides
作者: 林韋旭
Lin, Wei-Hsu
黃國華
洪耀欽
Huang, Gue-Wha Steven
Hung, Yao-Ching
材料科學與工程學系奈米科技碩博士班
關鍵字: 疫苗載體;奈米金粒子;免疫;禽流感病毒;口蹄疫病毒;Vaccine carrier;Gold nanoparticles;Immunoglobulin;H5N1;FMDV
公開日期: 2008
摘要: 本實驗室運用奈米金粒子作為高效率的疫苗載體,探討粒徑大小與抗體反應間的關係。首先將製備完成的口蹄疫病毒胜肽與禽流感病毒胜肽,與不同粒徑的奈米金(2奈米、5奈米、8奈米、12奈米、17奈米、37奈米至50奈米)做結合,為了確保能達到最大的鍵結量,因此我們在胜肽的酸基末端修飾一個半胱氨酸,強化鍵結能力。對照組方面,則以傳統的血氰蛋白與胜肽作結合之後,兩組接施打至老鼠體內去引發免疫反應。老鼠施打四周、六周、八周和十周後,開始採集血液,粒用免疫酵素檢測法,針對不同的胜肽與載體進行檢測。我們發現奈米金粒子在粒徑2奈米、5奈米、8奈米、12奈米和17奈米時,有良好的抗體反應。其最大的抗體反應量,其粒徑範圍是在8奈米至17奈米之間,與傳統的血氰蛋白的方式相比,其抗體反應量提高了將近三倍。值得注意的是,奈米金粒子載體本身並無檢測到任何抗體反應產生,但是血氰蛋白本身卻引起相當高的抗體反應,因此干擾了其所攜帶胜肽的抗體反應。此外,我們使用離子藕荷電漿分析儀計算分佈於老鼠脾臟中的奈米金粒子含量,我們發現此種免疫性質與不同粒徑的奈米金粒子在老鼠脾臟中的含量高度相關。在此研究中,我們發現奈米金粒子不但具有集中抗體辨識的功效,還可提高抗體反應的能力,並且奈米金粒子的粒徑大小與免疫行為有高度相關,其奈米金粒子粒徑範圍從8奈米至17奈米間,可當作最理想的胜肽疫苗載體。
To elicit the size-dependent carrier ability of gold nanoparticles (GNPs), synthetic peptide corresponding to foot-and-mouth disease virus (FMDV) and influenza A virus subtype (H5N1) viral proteins is conjugated to GNPs with diameter 2-nm, 5-nm, 8-nm, 12-nm, 17-nm, 37-nm, and 50-nm. Extra cysteine is added to the C-terminus of the FMDV peptide (pFMDV) and H5N1 peptide (pH5N1) to ensure maximum conjugation. Immunization of peptide-keyhole limpet hemocyanin (KLH) conjugate was performed as control. Blood are withdrawn on week 4, 6, 8, and 10. Titers against peptide and carriers are obtained. For peptide-GNP immunization, specific binding against peptide is detected in the sera of mice injected with 2-nm, 5-nm, 8-nm, 12-nm and 17-nm GNP conjugates. Maximum binding occur with GNPs of sizes between 8-nm to 17-nm. The peptide -GNPs induce 2.17 ~ 3 fold antibody responsecompared to peptide-KLH. In particular, all sera exhibited undetectable binding against GNP, while antisera of peptide-KLH present high levels of binding activity against KLH. Deposition of GNPs in mouse spleen was evaluated by ICP-MS. The immunogenicity of peptide is correlated to amounts of GNPs accumulated in spleen. In conclusion, we show the size-dependent immunogenic properties of peptide-GNP conjugates. GNPs ranging from 8-nm to 17-nm may serve as ideal carrier to elicit focused and enhanced antibody response against a synthetic peptide.
URI: http://140.113.39.130/cdrfb3/record/nctu/#GT079652518
http://hdl.handle.net/11536/43294
顯示於類別:畢業論文


文件中的檔案:

  1. 251801.pdf
  2. 251802.pdf
  3. 251803.pdf

若為 zip 檔案,請下載檔案解壓縮後,用瀏覽器開啟資料夾中的 index.html 瀏覽全文。