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Microarrays Incorporating Gold Grid Patterns for Protein Quantification.
ACS Omega ( IF 3.7 ) Pub Date : 2020-07-02 , DOI: 10.1021/acsomega.0c01549
Chang-Hyuk Yoo 1, 2 , Jae-Kyoung Yu 2 , Yeju Seong 2 , Jun-Kyu Choi 2
Affiliation  

Protein microarrays are miniaturized two-dimensional arrays, incorporating thousands of immobilized proteins, typically printed in minute amounts on functionalized solid substrates, which can be analyzed in a high-throughput fashion. Irreproducibility of the printing techniques adopted, resulting in inconsistently and nonuniformly deposited microscopic spots, nonuniform signal intensities from the printed microspots, and significantly high background noise are some of the critical issues that affect protein analysis using traditional protein microarrays. To overcome such issues, in this study, we introduced a novel gold grid pattern-based protein microarray. The grid patterns incorporated in our microarray are equivalent to the spots used for protein analysis in conventional protein microarrays. We utilized the signal intensities from the grid patterns acting as spots for quantifying the protein concentration levels. To demonstrate the utility of our novel design concept, we quantified as low as 66.7 ng/mL of bovine serum albumin using our gold grid pattern-based protein microarray. Our grid pattern-based design concept for protein quantification overcame the signal nonuniformity issues and ensured that the dominance of any distorted signal from a single spot did not affect the overall protein quantification results as encountered in conventional protein microarrays.

中文翻译:

结合金网格模式进行蛋白质定量的微阵列。

蛋白质微阵列是微型化的二维阵列,结合了成千上万种固定化蛋白质,通常以微量印刷在功能化固体基质上,可以以高通量方式进行分析。所采用的印刷技术的不可再现性,导致不一致和不均匀沉积的微观斑点,来自印刷的微斑点的信号强度不一致,以及显着高的背景噪声,这是影响使用传统蛋白质微阵列进行蛋白质分析的一些关键问题。为了克服这些问题,在这项研究中,我们介绍了一种新型的基于金网格模式的蛋白质微阵列。纳入我们的微阵列的网格图案等同于常规蛋白质微阵列中用于蛋白质分析的斑点。我们利用来自网格模式的信号强度作为斑点来量化蛋白质浓度水平。为了证明我们新颖的设计理念的实用性,我们使用基于金网格模式的蛋白质微阵列将牛血清白蛋白定量至66.7 ng / mL。我们基于网格模式的蛋白质定量设计理念克服了信号不均匀性问题,并确保了来自单个斑点的任何失真信号的优势都不会影响常规蛋白质微阵列所遇到的总体蛋白质定量结果。
更新日期:2020-07-14
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