Instrument-Free Protein Microarray Fabrication for Accurate Affinity Measurements.
Instrument-Free Protein Microarray Fabrication for Accurate Affinity Measurements.
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无需仪器的蛋白质微阵列制造，可实现准确的亲和力测量。
DOI：
10.3390/bios10110158
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发表时间：
2020-10-29
期刊：
影响因子：
--
通讯作者：
Ünlü S
中科院分区：
文献类型：
--
作者：
Celebi I;Geib MT;Chiodi E;Lortlar Ünlü N;Ekiz Kanik F;Ünlü S
蛋白质微阵列作为药物和生物标记物开发以及诊断等各个领域的有吸引力的工具而受到欢迎。因此，微阵列形式的多重结合亲和力测量变得至关重要。基于微阵列的蛋白质检测的制备依赖于探针溶液的精确分配以实现有效固定到活性表面上。设备成本过高，并且需要训练有素的人员来操作用于微阵列制造的高复杂性机器人点样器，这对研究人员，尤其是资源有限的小型实验室来说，是极大的损害。在这里，我们提出了一种低成本、无仪器的分配技术，熟悉微量移液的用户可以手动创建多重蛋白质检测，与机器人点样检测相比，该检测显示出更高的捕获效率和噪音水平。在本研究中，我们使用干涉反射成像传感器平台，通过分析与抗 α-乳清蛋白抗体相互作用获得的结合动力学，比较了手动和自动分配 α-乳清蛋白探针点的效率。我们证明，通过微量移液器手动点样制备的蛋白质阵列达到并超过了最先进的机器人点样器制备的蛋白质阵列的性能。与通过平均对应相同有效传感器表面积的 75 个机器人点获得的数据相比，这些无需仪器的蛋白质检测具有更高的结合信号（约 4 倍的改进）和约 3 倍的结合曲线信噪比 (SNR)。我们展示了以 24 路多重芯片格式确定抗原抗体结合系数的潜力，且测量误差小于 5%。
Protein microarrays have gained popularity as an attractive tool for various fields, including drug and biomarker development, and diagnostics. Thus, multiplexed binding affinity measurements in microarray format has become crucial. The preparation of microarray-based protein assays relies on precise dispensing of probe solutions to achieve efficient immobilization onto an active surface. The prohibitively high cost of equipment and the need for trained personnel to operate high complexity robotic spotters for microarray fabrication are significant detriments for researchers, especially for small laboratories with limited resources. Here, we present a low-cost, instrument-free dispensing technique by which users who are familiar with micropipetting can manually create multiplexed protein assays that show improved capture efficiency and noise level in comparison to that of the robotically spotted assays. In this study, we compare the efficiency of manually and robotically dispensed α-lactalbumin probe spots by analyzing the binding kinetics obtained from the interaction with anti-α-lactalbumin antibodies, using the interferometric reflectance imaging sensor platform. We show that the protein arrays prepared by micropipette manual spotting meet and exceed the performance of those prepared by state-of-the-art robotic spotters. These instrument-free protein assays have a higher binding signal (~4-fold improvement) and a ~3-fold better signal-to-noise ratio (SNR) in binding curves, when compared to the data acquired by averaging 75 robotic spots corresponding to the same effective sensor surface area. We demonstrate the potential of determining antigen-antibody binding coefficients in a 24-multiplexed chip format with less than 5% measurement error.
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DOI：
10.1007/978-1-4939-9616-2_2
发表时间：
2019-01-01
期刊：
BIOMIMETIC SENSING
影响因子：
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作者：
Needham, James W.;Unlu, Nese Lortlar;Unlu, M. Selim
通讯作者：
Unlu, M. Selim
影响因子：
7.4
作者：
Pirri, G;Damin, F;Depero, LE
通讯作者：
Depero, LE
影响因子：
6.1
作者：
Finetti, Chiara;Plavisch, Lauren;Chiari, Marcella
通讯作者：
Chiari, Marcella
影响因子：
3.3
作者：
Couture, Maxime;Zhao, Sandy Shuo;Masson, Jean-Francois
通讯作者：
Masson, Jean-Francois
干涉反射成像传感器（IRIS） - 一种用于多重诊断和数字检测的平台技术。
DOI：
10.3390/s150717649
发表时间：
2015-07-20
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
Avci O;Ünlü NL;Özkumur AY;Ünlü MS
通讯作者：
Ünlü MS