Surface Functionalization Methods to Enhance Bioconjugation in Metal-Labeled Polystyrene Particles.
Surface Functionalization Methods to Enhance Bioconjugation in Metal-Labeled Polystyrene Particles.
复制标题
DOI：
10.1021/ma200582q
复制
发表时间：
2011-06-28
期刊：
影响因子：
5.5
通讯作者：
Winnik MA
中科院分区：
文献类型：
--
作者：
Abdelrahman AI;Thickett SC;Liang Y;Ornatsky O;Baranov V;Winnik MA
通过分散聚合合成的镧系编码聚苯乙烯颗粒是基于细胞计数的免疫分析的优秀候选物，然而，它们以前缺乏将生物分子偶联到颗粒表面的能力。我们在这里提出了三种方法来后功能化这些颗粒，使蛋白质的共价附着。我们的第一种方法是使用部分水解的聚（n -乙烯基吡咯烷酮）作为分散聚合稳定剂来合成颗粒表面具有高浓度-COOH基团的颗粒。在另一种为镧系编码粒子提供-COOH功能的策略中，我们采用种子乳液聚合将聚甲基丙烯酸（PMAA）链接枝到这些粒子的表面。然而，这两种方法在生物偶联的程度上几乎没有改善。在我们的第三种方法中，种子乳液聚合随后被用作一种方法，在这些颗粒的表面上生长功能聚合物外壳（在这种情况下，聚甲基丙烯酸甘油酯（PGMA）），这被证明是非常成功的。富含环氧化物的PGMA外壳允许NeutrAvidin广泛的表面生物偶联，通过lu标记的生物素报告器进行探测（每颗粒约7 × 105个结合事件，非特异性结合量非常低），并通过大量细胞术进行分析。结果表明，不需要偶联剂如EDC，这是由于颗粒表面的反应性。这些粒子是稳定的，聚合物外壳的加入并不影响粒子内部狭窄的镧系离子分布。这些粒子代表了迄今为止基于镧系元素标记粒子的高度多重生物测定的最有希望的候选物。
Lanthanide-encoded polystyrene particles synthesized by dispersion polymerization are excellent candidates for mass cytometry based immunoassays, however they have previously lacked the ability to conjugate biomolecules to the particle surface. We present here three approaches to post-functionalize these particles, enabling the covalent attachment of proteins. Our first approach used partially hydrolyzed poly(N-vinylpyrrolidone) as a dispersion polymerization stabilizer to synthesize particles with high concentration of -COOH groups on the particle surface. In an alternative strategy to provide -COOH functionality to the lanthanide-encoded particles, we employed seeded emulsion polymerization to graft poly(methacrylic acid) (PMAA) chains onto the surface of these particles. However, these two approaches gave little to no improvement in the extent of bioconjugation. In our third approach, seeded emulsion polymerization was subsequently used as a method to grow a functional polymer shell (in this case, poly(glycidyl methacrylate) (PGMA)) onto the surface of these particles, which proved highly successful. The epoxide-rich PGMA shell permitted extensive surface bioconjugation of NeutrAvidin, as probed by an Lu-labeled biotin reporter (ca. 7 × 105 binding events per particle with a very low amount of non-specific binding) and analyzed by mass cytometry. It was shown that coupling agents such as EDC were not needed, such was the reactivity of the particle surface. These particles were stable and the addition of a polymeric shell was shown did not affect the narrow lanthanide ion distribution within the particle interior as analyzed by mass cytometry. These particles represent the most promising candidates for the development of a highly multiplexed bioassay based on lanthanide-labeled particles to date.
登录
查看更多内容
影响因子：
7.4
作者：
GRABAR, KC;FREEMAN, RG;NATAN, MJ
通讯作者：
NATAN, MJ
影响因子：
2.9
作者：
Horák, D;Karpísek, M;Benes, M
通讯作者：
Benes, M
影响因子：
2.9
作者：
Lathia US;Ornatsky O;Baranov V;Nitz M
通讯作者：
Nitz M
影响因子：
56.9
作者：
Bruchez, M;Moronne, M;Alivisatos, AP
通讯作者：
Alivisatos, AP
影响因子：
9.9
作者：
Kawaguchi, S;Yekta, A;Winnik, MA
通讯作者：
Winnik, MA