Department of Biomedical Engineering, Pratt School of Engineering , Duke University , Durham , North Carolina 27708 , United States.
Langmuir. 2019 Feb 5;35(5):1379-1390. doi: 10.1021/acs.langmuir.8b01597. Epub 2018 Aug 21.
Motivated by the lack of adventitious protein adsorption on zwitterionic polymer brushes that promise low noise and hence high analytical sensitivity for surface-based immunoassays, we explored their use as a substrate for immunoassay fabrication by the inkjet printing of antibodies. We observed that a poly(sulfobetaine)methacrylate brush on glass is far too hydrophilic to enable the noncovalent immobilization of antibodies by inkjet printing. To circumvent this limitation, we developed a series of hybrid zwitterionic-cationic surface coatings with tunable surface wettability that are suitable for the inkjet printing of antibodies but also have low protein adsorption. We show that in a microarray format in which both the capture and detection antibodies are discretely printed as spots on these hybrid brushes, a point-of-care sandwich immunoassay can be carried out with an analytical sensitivity and dynamic range that is similar to or better than those of the same assay fabricated on a PEG-like brush. We also show that the hybrid polymer brushes do not bind anti-PEG antibodies that are ubiquitous in human blood, which can be a problem with immunoassays fabricated on PEG-like coatings.
受两性离子聚合物刷在表面基免疫分析中具有低噪声、高分析灵敏度的承诺的启发,我们探索了将其用作喷墨打印抗体的免疫分析制备的基底的用途。我们观察到,玻璃上的聚(磺基甜菜碱)甲基丙烯酸酯刷具有极高的亲水性,以至于无法通过喷墨打印实现抗体的非共价固定化。为了克服这一限制,我们开发了一系列具有可调润湿性的混合两性离子-阳离子表面涂层,这些涂层适合喷墨打印抗体,但蛋白质吸附性低。我们表明,在微阵列格式中,捕获抗体和检测抗体都以离散的斑点形式打印在这些混合刷上,可以进行即时点的夹心免疫分析,其分析灵敏度和动态范围与在类似 PEG 的刷上制备的相同测定法相似或更好。我们还表明,混合聚合物刷不与在人血液中普遍存在的抗聚乙二醇抗体结合,这在类似 PEG 的涂层上制备的免疫分析中可能是一个问题。