Lanthanide-containing polymer microspheres by multiple-stage dispersion polymerization for highly multiplexed bioassays.
Lanthanide-containing polymer microspheres by multiple-stage dispersion polymerization for highly multiplexed bioassays.
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DOI：
10.1021/ja9052009
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发表时间：
2009-10-28
影响因子：
15
通讯作者：
Winnik, Mitchell A.
中科院分区：
文献类型：
--
作者：
Abdelrahman, Ahmed I.;Dai, Sheng;Thickett, Stuart C.;Ornatsky, Olga;Bandura, Dmitry;Baranov, Vladimir;Winnik, Mitchell A.
我们描述了金属编码聚苯乙烯微球的合成和表征，通过多级分散聚合，直径约为2 μm，尺寸分布非常窄。在苯乙烯转化率约10%后，通过向反应中添加溶解在乙醇中的LnCl 3盐和过量丙烯酸或甲基丙烯酸乙酰乙酰乙酯（AAEM）的混合物，将不同的镧系元素负载到这些微球中，即，在颗粒成核阶段完成之后很久。单个微球含有ca. 106 - 108单一元素或元素混合物的螯合镧系元素离子。这些微球的特点是一个接一个，利用一种新的质谱仪与电感耦合等离子体（ICP）电离源和飞行时间（TOF）质谱检测。合成了含有不同浓度水平的一系列不同金属的微球，以满足二进制编码和枚举编码协议的要求。用四种不同的金属在五个浓度水平下，我们可以实现624的变异性，我们报告的策略应该允许获得更大的变异性。为了证明元件编码的珠粒对于高度多重免疫测定的有用性，我们进行了原理验证模型生物测定，其涉及将小鼠IgG缀合至含有La和Tm的颗粒的表面，并通过带有金属螯合聚合物的抗小鼠IgG与Pr进行检测。
We describe the synthesis and characterization of metal-encoded polystyrene microspheres by multiple-stage dispersion polymerization with diameters on the order of 2 µm and a very narrow size distribution. Different lanthanides were loaded into these microspheres through the addition of a mixture of LnCl3 salts and excess acrylic acid or acetoacetylethyl methacrylate (AAEM) dissolved in ethanol to the reaction after about 10% conversion of styrene, i.e., well after the particle nucleation stage was complete. Individual microspheres contain ca. 106 – 108 chelated lanthanide ions, of either a single element or a mixture of elements. These microspheres were characterized one-by-one utilizing a novel mass cytometer with an inductively coupled plasma (ICP) ionization source and time-of-flight (TOF) mass spectrometry detection. Microspheres containing a range of different metals at different levels of concentration were synthesized to meet the requirements of binary encoding and enumeration encoding protocols. With four different metals at five levels of concentration, we could achieve a variability of 624, and the strategy we report should allow one to obtain much larger variability. To demonstrate the usefulness of element-encoded beads for highly multiplexed immunoassays, we carried out a proof-of-principle model bioassay involving conjugation of mouse IgG to the surface of La and Tm containing particles, and its detection by an anti-mouse IgG bearing a metal-chelating polymer with Pr.
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影响因子：
15
作者：
Song, JS;Tronc, F;Winnik, MA
通讯作者：
Winnik, MA
影响因子：
2.3
作者：
Paschkunova-Martic, I;Kremser, C;Debbage, P
通讯作者：
Debbage, P
影响因子：
7.4
作者：
Matsuya, T;Tashiro, S;Kataoka, K
通讯作者：
Kataoka, K
影响因子：
8.6
作者：
Morgan, E;Varro, R;Gaur, A
通讯作者：
Gaur, A
影响因子：
2.2
作者：
Cook, EB;Stahl, JL;Barney, NP
通讯作者：
Barney, NP