Characterizing the Incorporation of DNA into Single NIPAm Hydrogel Nanoparticles with Surface Plasmon Resonance Imaging Measurements
Characterizing the Incorporation of DNA into Single NIPAm Hydrogel Nanoparticles with Surface Plasmon Resonance Imaging Measurements
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DOI：
10.1021/acs.jpcc.9b00444
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发表时间：
2019-02
期刊：
影响因子：
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通讯作者：
B. Matthews;A. Maley;Kellen M. Kartub;R. Corn
中科院分区：
文献类型：
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作者：
B. Matthews;A. Maley;Kellen M. Kartub;R. Corn
合成了掺入30 mer单链DNA（ssDNA）寡核苷酸的基于N-异丙基丙烯酰胺的水凝胶纳米颗粒（HNP），并用单纳米颗粒表面等离子体共振成像（SPRI）显微镜、动态光散射、透射电子显微镜和荧光测量进行表征。合成的HNP具有230 nm的平均直径，并且与没有DNA的HNP相比，表现出平均单纳米颗粒SPRI折射率（Δ% RNP）的大的（5-10倍）增加。使用SPRI和荧光测量的组合来测量约20 000个互补ssDNA到每个HNP中的吸收，得到4.89 × 108 M-1的Langmuir等温吸附系数。单纳米颗粒SPRI测量还显示，约35%的掺入的ssDNA可用于杂交摄取和使用核酸外切酶I的酶水解。我们将纳米颗粒中无活性ssDNA群体的存在归因于一种复合物。
N-Isopropylacrylamide-based hydrogel nanoparticles (HNPs) that incorporate 30 mer single-stranded DNA (ssDNA) oligonucleotides were synthesized and characterized with single-nanoparticle surface plasmon resonance imaging (SPRI) microscopy, dynamic light scattering, transmission electron microscopy, and fluorescence measurements. The synthesized HNPs had an averaged diameter of 230 nm and exhibited a large (5–10×) increase in the average single-nanoparticle SPRI refractive index (Δ% RNP), as compared with HNPs without DNA. A combination of SPRI and fluorescence measurements was used to measure the uptake of approximately 20 000 complementary ssDNA into each HNP, resulting in a Langmuir isotherm adsorption coefficient of 4.89 × 108 M–1. Single-nanoparticle SPRI measurements also showed that approximately 35% of the incorporated ssDNA was accessible for both hybridization uptake and enzymatic hydrolysis using exonuclease I. We attribute the presence of an inactive ssDNA population in the nanoparticle to a co...