螢光粒子在生化分析上的初步應用

Preliminary Study of Fluorescent Particles for Biochemical Analysis

李怡青(Yi-Ching Lee)

指導教授 ： 傅傳博

國立暨南國際大學/科技學院/應用化學系/碩士(2008年)

摘要

本論文係關於製備功能性奈米粒子( functional nanoparticles )並結合免疫吸附分析法 ( immunosorbent assay )做生化分離上之應用研究。
本研究成功製備40nm、80 nm 及150nm之 Fe3O4 粒子。磁性粒子表面經有機物質修飾粒子表面後粒徑為80 nm、150 nm、200nm。
接下來，將鏈黴抗生素蛋白 (Streptavidin) 或免疫球蛋白 (IgG) 固定化在 SiO2 包覆四氧化三鐵磁性載體上，再將生物素 (d-Biotin) 或蛋白質A (Protein A) 固定在帶有螢光染料的載體上，結合微盤做簡單生化分析上之測試研究。初步探討磁性載體大小，及顆粒數目在生化分析上的可能應用。
此次研究主要是運用鏈黴抗生物素蛋白（Streptavidin）－生物素（Biotin）和免疫球蛋白G（IgG）－蛋白質A（Protein A）這兩對抗原與抗體之間在顆粒間的作用。實驗設計中，我們改變顆粒大小、及注入顆粒數的不同等條件來進行沉積試驗。在磁場的作用下，由於他們具有很強的專一性，帶有磁性顆粒的Streptavidin 和IgG 可以成功地分別與標示有Biotin 和Protein A 之螢光顆粒二氧化矽顆粒作用，可作進一步的分析。在控制組實驗的對照之下，充分表現出好的專一性與高度的選擇性。

關鍵字

功能性奈米粒子

並列摘要

The goal of this study is to synthesize functional magnetic particles and combine with immunosorbent assay for biochemical analysis.
The sizes of magnetic nanoparticles were 40nm, 80 nm and 150nm, and SiO2 coated Fe3O4 were 80nm, 150 nm and 200nm.
Streptavidin or IgG was bound onto Fe3O4@SiO2 particles. Biotin or protein A was covalently bound to fluorescent marker, and applied to magnetic separation in microplate. We studied experimental parameters of magnetic particles size、and particles number for immunoassay. Preliminary study of immunoassay between particles was studied to demonstrate the potential application of this method for biochemical analysis.
The interactions of streptavidin-biotin and IgG-protein A that coating on particles was studied. Experimental parameters were studied under different conditions of the particle size and the number of particles. The streptavidin-conjugated magnetic
beads and IgG-conjugated magnetic beads can capture fluorescent silica particles labeled with biotin and protein A in the 96 well plate under magnetic filed. The selectivity is high with controlled experiments.

並列關鍵字

functional nanoparticles

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