Hong Woogyeong, Jeong Seong-Geun, Shim Gyurak, Kim Dae Young, Pack Seung Pil, Lee Chang-Soo
1Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Seoul, Korea.
2New Drug Development Center, Osong Medical Innovation Foundation, Seoul, Korea.
Biotechnol Bioprocess Eng. 2018;23(6):686-692. doi: 10.1007/s12257-018-0430-2. Epub 2019 Jan 17.
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.
纸质分析装置（PADs）因其价格低廉且便于携带，已在许多领域得到广泛应用。对于可重复的定量分析而言，将蛋白质牢固地固定在PADs上至关重要。传统的将蛋白质固定在PADs上的技术基于物理吸附，但蛋白质很容易因微弱的物理力而被去除。因此，使用物理吸附难以确保PADs分析结果的可重复性。为克服这一局限性，在本研究中，我们展示了一种将蛋白质与纤维素纸共价结合的方法。该方法包括三个步骤，即纸张的高碘酸盐氧化、席夫碱的形成和还原胺化。我们使用傅里叶变换红外光谱（FT-IR）分析鉴定了纸张上形成的醛基和亚胺基。这种共价结合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法确认了固定化抗体的活性。我们期望这种固定化方法将有助于PADs以高重现性和灵敏度实现商业化。
