Highly efficient binding of paramagnetic beads bioconjugated with 100,000 or more antibodies to protein-coated surfaces.
Highly efficient binding of paramagnetic beads bioconjugated with 100,000 or more antibodies to protein-coated surfaces.
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DOI：
10.1021/ac3028257
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发表时间：
2012-12-04
影响因子：
7.4
通讯作者：
Rusling, James F.
中科院分区：
文献类型：
--
作者：
Mani, Vigneshwaran;Wasalathanthri, Dhanuka P.;Joshi, Arnit A.;Kumar, Challa V.;Rusling, James F.
我们在此报告了 1 μm 超直径顺磁颗粒 (MP) 的首次动力学表征，该颗粒装饰有超过 100,000 个与附着在平坦表面上的蛋白质抗原结合的抗体。表面等离子共振 (SPR) 用于表明这些抗体衍生的 MP (MP-Ab2) 表现出不可逆的结合，与游离抗体相比，结合率增加了 100 倍。 MP-Ab2 从 SPR 传感器表面的解离常数的估计上限为 5 fM，而游离抗体的解离常数上限为 3-8 nM。这些结果可以通过 MP-Ab2 与蛋白质修饰表面多达 2000 次的相互作用来解释。研究结果与 MP-Ab2 在溶液中高效捕获蛋白质抗原相一致，并部分解释了这些珠子在 fM 和 aM 范围内超灵敏蛋白质检测中的用途。这些颗粒在 SPR 芯片上的聚集（可能是由于颗粒中残留的磁性微区）也有助于超灵敏检测，并且还可能有助于驱动不可逆结合。
We report here the first kinetic characterization of 1 μm super diameter paramagnetic particles (MP) decorated with over 100,000 antibodies binding to protein antigens attached to flat surfaces. Surface plasmon resonance (SPR) was used to show that these antibody-derivatized MPs (MP-Ab2) exhibit irreversible binding with 100-fold increased association rates compared to free antibodies. The estimated upper limit for the dissociation constant of MP-Ab2 from the SPR sensor surface is 5 fM, compared to 3–8 nM for the free antibodies. These results are explained by up to 2000 interactions of MP-Ab2 with protein-decorated surfaces. Findings are consistent with highly efficient capture of protein antigens in solution by the MP-Ab2, and explain in part the utility of these beads for ultrasensitive protein detection into the fM and aM range. Aggregation of these particles on the SPR chip, probably due to residual magnetic microdomains in the particles, also contributes to ultrasensitive detection and may also help drive the irreversible binding.
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影响因子：
12.6
作者：
Chikkaveeraiah, Bhaskara V.;Mani, Vigneshwaran;Patel, Vyomesh;Gutkind, J. Silvio;Rusling, James F.
通讯作者：
Rusling, James F.
影响因子：
4.9
作者：
Lin, CC;Yeh, YC;Chen, CC
通讯作者：
Chen, CC
影响因子：
16.6
作者：
Krishnan, Sadagopan;Mani, Vigneshwaran;Wasalathanthri, Dhanuka;Kumar, Challa V.;Rusling, James F.
通讯作者：
Rusling, James F.
DOI：
10.1039/c0an00204f
发表时间：
2010-10
期刊：
The Analyst
影响因子：
--
作者：
Rusling JF;Kumar CV;Gutkind JS;Patel V
通讯作者：
Patel V
影响因子：
3.4
作者：
Schuck, P
通讯作者：
Schuck, P