Photo- and biophysical studies of lectin-conjugated fluorescent nanoparticles: reduced sensitivity in high density assays.
Photo- and biophysical studies of lectin-conjugated fluorescent nanoparticles: reduced sensitivity in high density assays.
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DOI：
10.1021/jp101854m
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发表时间：
2010-11-18
影响因子：
3.3
通讯作者：
Zimmt, Matthew B.
中科院分区：
文献类型：
--
作者：
Wang, Yaqi;Gildersleeve, Jeffrey C.;Basu, Amit;Zimmt, Matthew B.
已经开发了凝集素缀合的荧光二氧化硅纳米颗粒（fNP）用于上皮组织活检的基于碳水化合物的组织病理学评价。选择fNP平台是因为其与直接染料标记相比增强的发射亮度。进行碳水化合物微阵列研究以比较甘露糖识别凝集素伴刀豆球蛋白A（ConA）在缀合至荧光二氧化硅纳米颗粒（ConA-fNP）之前和之后的碳水化合物选择性。这些研究揭示了与生物素-ConA/Cy 3-链霉亲和素染色相比，用ConA-fNP染色后令人惊讶的低发射强度。进行fNP和ConA-fNP缀合物的一系列生物物理和生物物理表征以探测微阵列测定中来自fNP的低灵敏度。将多达1200个荧光素（FL）和80个四甲基罗丹明（TR）染料分子掺入46 nm直径的fNP中，分别产生比单个染料分子大400倍和35倍的发射亮度。与羧酸表面修饰的纳米颗粒缀合的ConA凝集素覆盖fNP表面的15-30%。与可溶性ConA相比，与fNP缀合的ConA的CD光谱和甘露糖底物选择性略有不同。尽管fNP的高发射亮度增强了具有低分析物密度的样品的检测灵敏度，但是大的fNP直径限制了fNP募集和与具有高分析物密度的样品的结合。碳水化合物微阵列的高分析物密度和接近二维的目标格式使得探针尺寸成为关键参数。在该应用中，与直接染料标记相比，fNP标记提供最小的灵敏度优势。
Lectin conjugated, fluorescent silica nanoparticles (fNP) have been developed for carbohydrate based histopathology evaluations of epithelial tissue biopsies. The fNP platform was selected for its enhanced emissive brightness compared to direct dye labeling. Carbohydrate microarray studies were performed to compare the carbohydrate selectivity of the mannose recognizing lectin Concanavalin A (ConA) before and after conjugation to fluorescent silica nanoparticles (ConA-fNP). These studies revealed surprisingly low emission intensities upon staining with ConA-fNP compared to biotin-ConA / Cy3-strepavidin staining. A series of photophysical and biophysical characterizations of the fNP and ConA-fNP conjugates were performed to probe the low sensitivity from fNP in the microarray assays. Up to 1200 fluorescein (FL) and 80 tetramethylrhodamine (TR) dye molecules were incorporated into 46 nm diameter fNP, yielding emissive brightness’ 400 and 35 times larger than the individual dye molecules, respectively. ConA lectin conjugated to carboxylic acid surface modified nanoparticles covers 15–30% of the fNP surface. The CD spectra and mannose substrate selectivity of ConA conjugated to the fNP differed slightly compared to soluble ConA. Although, the high emissive brightness of fNP enhances detection sensitivity for samples with low analyte densities, large fNP diameters limit fNP recruitment and binding to samples with high analyte densities. The high analyte density and nearly two-dimensional target format of carbohydrate microarrays make probe size a critical parameter. In this application, fNP labels afford minimal sensitivity advantage compared to direct dye labeling.
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影响因子：
--
作者：
ISAK, SJ;EYRING, EM
通讯作者：
EYRING, EM
DOI：
10.1073/pnas.86.7.2190
发表时间：
1989-04-01
影响因子：
11.1
作者：
HENDRICKSON, WA;PAHLER, A;PHIZACKERLEY, RP
通讯作者：
PHIZACKERLEY, RP
影响因子：
4.4
作者：
Lelpold, Mtchael D.;Herrera, Isaac;Ornatsky, Olga;Baranov, Vladimir;Nitz, Mark
通讯作者：
Nitz, Mark
影响因子：
1.9
作者：
Hill, SC;Pinnick, RG;Chang, RK
通讯作者：
Chang, RK
影响因子：
14.8
作者：
Hsu, Ku-Lung;Mahal, Lara K.
通讯作者：
Mahal, Lara K.