Improvement in the Reproducibility of a Paper-based Analytical Device (PAD) Using Stable Covalent Binding between Proteins and Cellulose Paper
Improvement in the Reproducibility of a Paper-based Analytical Device (PAD) Using Stable Covalent Binding between Proteins and Cellulose Paper
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DOI：
10.1007/s12257-018-0430-2
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发表时间：
2018-11-01
影响因子：
3.2
通讯作者：
Lee, Chang-Soo
中科院分区：
文献类型：
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作者：
Hong, Woogyeong;Jeong, Seong-Geun;Lee, Chang-Soo
纸基分析仪器（PAD）由于价格实惠、便于携带，在许多领域得到了广泛应用。为了进行可重复的定量分析，在PAD上强烈标记蛋白质是至关重要的。用于将蛋白质固定在PAD上的常规技术是基于物理吸附，但是蛋白质可以通过弱的物理力容易地去除。因此，很难确保使用物理吸附的PAD的分析结果的再现性。为了克服这一局限性，在这项研究中，我们展示了一种将蛋白质共价结合到纤维素纸上的方法。该方法由三个步骤组成，包括纸的高碘酸盐氧化、席夫碱的形成和还原胺化。我们确定了醛和亚胺基团形成的纸上使用FT-IR分析。这种共价键方法增强了蛋白质的结合力和结合能力。我们通过夹心免疫测定证实了固定化抗体的活性。我们希望这种固定化方法将有助于商业化的PAD具有高重现性和灵敏度。
Paper-based analytical devices (PADs) have been widely used in many fields because they are affordable and portable. For reproducible quantitative analysis, it is crucial to strongly immobilize proteins on PADs. Conventional techniques for immobilizing proteins on PADs are based on physical adsorption, but proteins can be easily removed by weak physical forces. Therefore, it is difficult to ensure the reproducibility of the analytical results of PADs using physical adsorption. To overcome this limitation, in this study, we showed a method of covalent binding of proteins to cellulose paper. This method consists of three steps, which include periodate oxidation of paper, the formation of a Schiff base, and reductive amination. We identified aldehyde and imine groups formed on paper using FT-IR analysis. This covalent bonding approach enhanced the binding force and binding capacity of proteins. We confirmed the activity of an immobilized antibody through a sandwich immunoassay. We expect that this immobilization method will contribute to the commercialization of PADs with high reproducibility and sensitivity.