Kurihara Yoshikazu, Yokota Hiroyuki, Takahashi Masaru
Konica Minolta, Inc., 1 Sakura-machi, Hino-shi, Tokyo 191-8511, Japan.
ACS Omega. 2022 Nov 3;7(45):41641-41650. doi: 10.1021/acsomega.2c05653. eCollection 2022 Nov 15.
In this study, we developed a simple method for preparing highly dispersed, stable, and streptavidin (SA)-functionalized carboxymethyl dextran (CMD)-coated melamine nanoparticles (MNPs) in an aqueous buffer at neutral pH. Dynamic light scattering (DLS) revealed the agglomeration of MNPs in an aqueous buffer at neutral pH. When CMD, -hydroxysuccinimide (NHS), and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) were simultaneously mixed with the MNPs, CMD was bound to the MNPs, promoting their dispersibility. Preparation of SA-CMD-MNPs was accomplished simply by adding SA solution to the CMD-MNPs. The amount of SA bound to the CMD-MNPs was quantified by the bicinchoninic assay, and the amount of SA molecules bound to each CMD-MNP was 417 ± 4. SA-CMD-MNPs exhibited high dispersity (polydispersity index = 0.058) in a neutral phosphate buffer and maintained it for 182 days with dispersion using a probe sonicator (5 s) before DLS characterization. The performance of the SA-CMD-MNPs in biosensing was evaluated by immunohistochemistry, which revealed that the nanoparticles could specifically stain MCF-7 cells derived from breast cancer cells with low HER2 expression. This study provides an effective method for synthesizing highly dispersible nanoparticles for biosensing.
在本研究中,我们开发了一种简单的方法,可在中性pH值的水性缓冲液中制备高度分散、稳定且具有链霉亲和素(SA)功能化的羧甲基葡聚糖(CMD)包覆的三聚氰胺纳米颗粒(MNP)。动态光散射(DLS)显示中性pH值的水性缓冲液中MNP会发生团聚。当CMD、N-羟基琥珀酰亚胺(NHS)和1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)与MNP同时混合时,CMD会与MNP结合,促进其分散性。SA-CMD-MNP的制备只需将SA溶液加入到CMD-MNP中即可完成。通过二辛可宁酸测定法定量分析与CMD-MNP结合的SA量,每个CMD-MNP结合的SA分子数量为417±4。SA-CMD-MNP在中性磷酸盐缓冲液中表现出高分散性(多分散指数=0.058),并且在进行DLS表征前使用探针超声仪分散(5秒)的情况下可保持182天。通过免疫组织化学评估了SA-CMD-MNP在生物传感中的性能,结果表明这些纳米颗粒可以特异性地对源自HER2低表达乳腺癌细胞的MCF-7细胞进行染色。本研究为合成用于生物传感的高度可分散纳米颗粒提供了一种有效方法。