Lee Seung-Woo, Ahn Junhyoung, Kim Min-Gon, Shin Yong-Beom, Lee Jae Jong, Lim Ki-Pil, Kim Ki-Bum
Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Korea.
J Nanosci Nanotechnol. 2010 May;10(5):3246-9. doi: 10.1166/jnn.2010.2266.
An enzyme-catalyzed precipitation reaction was employed as a means to increase the change in the LSPR signal after intermolecular bindings between antigens and antibodies occurred on gold nanodot surfaces. The gold nanodot array with an diameter of 175 nm and a thickness of 20 nm was fabricated on a glass wafer using thermal nanoimprint lithography. The human interleukin (hIL) 5 antibody was immobilized on the gold nanodot, followed by binding of hIL 5 to the anti-hIL 5. Subsequently, a biotinylated anti-hIL 5 and a alkaline phosphatase conjugated with streptavidin were simultaneously introduced. A mixture of 5-bromo-4-chloro-3-indolyl phosphate p-toluidine (BCIP) and nitro blue tetrazolium (NBT) was then used for precipitation, which resulted from the biocatalytic reaction of the alkaline phosphatase on gold nanodot. The LSPR spectra were obtained after each binding process. Using this analysis, the enzyme-catalyzed precipitation reaction on gold nanodots was found to be effective in amplifying the change in the peak wavelength of LSPR after molecular bindings.
采用酶催化沉淀反应，作为在金纳米点表面发生抗原与抗体分子间结合后增加局域表面等离子体共振（LSPR）信号变化的一种手段。使用热纳米压印光刻技术在玻璃晶片上制备了直径为175 nm、厚度为20 nm的金纳米点阵列。将人白细胞介素（hIL）5抗体固定在金纳米点上，随后hIL 5与抗hIL 5结合。随后，同时引入生物素化的抗hIL 5和与链霉亲和素缀合的碱性磷酸酶。然后使用5-溴-4-氯-3-吲哚磷酸对甲苯胺（BCIP）和硝基蓝四唑（NBT）的混合物进行沉淀，这是碱性磷酸酶在金纳米点上的生物催化反应导致的。在每个结合过程之后获得LSPR光谱。通过该分析发现，金纳米点上的酶催化沉淀反应在放大分子结合后LSPR峰值波长的变化方面是有效的。
