Emergent Properties of Nanosensor Arrays: Applications for Monitoring IgG Affinity Distributions, Weakly Affined Hypermannosylation, and Colony Selection for Biomanufacturing
Emergent Properties of Nanosensor Arrays: Applications for Monitoring IgG Affinity Distributions, Weakly Affined Hypermannosylation, and Colony Selection for Biomanufacturing
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DOI：
10.1021/nn403215e
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发表时间：
2013-09-01
期刊：
影响因子：
17.1
通讯作者：
Strano, Michael S.
中科院分区：
文献类型：
--
作者：
Reuel, Nigel F.;Grassbaugh, Brittany;Strano, Michael S.
广泛认识到，可寻址传感器阵列可以被多路复用以用于分析物文库的无标记检测。然而，这样的阵列具有有用的性质，从合奏出现，即使当单官能化。作为例子，我们表明，纳米传感器阵列可以估计所观察到的解离常数（K-D）的平均值和方差，使用三个不同的结合IgG与蛋白A作为识别位点的例子，包括人IgG（K-D mu = 19 μ M，sigma（2）= 1000 mM（2））、鼠IgG（K-D mu = 4.3 nM，sigma（2）= 3 μ M-2）和来自CHO细胞的人IgG（K-D mu = 2.5 nM，sigma（2）= 0.01 μ M-2）。第二，我们表明，纳米传感器阵列可以唯一地监测弱亲和分析物的相互作用，通过观察到的相互作用的数量增加。一种应用涉及使用PSA-凝集素缀合的传感器阵列监测来自CHO的人IgG的代谢诱导的高甘露糖基化，其中测量和比较时间糖基化模式。最后，传感器阵列还可以在空间上映射来自细胞生物合成的分析物的局部产生。作为一个例子，我们对直接在纳米传感器阵列本身上培养的产生IgG的HEK菌落的生产率进行了排名。
It is widely recognized that an array of addressable sensors can be multiplexed for the label-free detection of a library of analytes. However, such arrays have useful properties that emerge from the ensemble, even when monofunctionalized. As examples, we show that an array of nanosensors can estimate the mean and variance of the observed dissodation constant (K-D), using three different examples of binding IgG with Protein A as the recognition site, including human IgG (K-D mu = 19 mu M, sigma(2) = 1000 mM(2)), murine IgG (K-D mu = 4.3 nM, sigma(2) = 3 mu M-2), and human IgG from CHO cells (K-D mu = 2.5 nM, sigma(2) = 0.01 mu M-2). Second, we show that an array of nanosensors can uniquely monitor weakly affined analyte interactions via the Increased number of observed interactions. One application involves monitoring the metabolically induced hypermannosylation of human IgG from CHO using PSA-lectin conjugated sensor arrays where temporal glycosylation patterns are measured and compared. Finally, the array of sensors can also spatially map the local production of an analyte from cellular biosynthesis. As an example, we rank productivity of IgG-producing HEK colonies cultured directly on the array of nanosensors itself.