Department of Chemistry and Physics of Surfaces and Biointerfaces, Institute of Macromolecular Chemistry ASCR , v.v.i., Prague, Czech Republic.
Department of Chemistry, Doane University, Crete, Nebraska, and the Center for Nanohybrid Functional Materials (CNFM), University of Nebraska-Lincoln , Lincoln, Nebraska 68588, United States.
ACS Appl Mater Interfaces. 2017 Apr 5;9(13):12109-12117. doi: 10.1021/acsami.7b01184. Epub 2017 Mar 23.
Protein-repellent reactive surfaces that promote localized specific binding are highly desirable for applications in the biomedical field. Nonspecific adhesion will compromise the function of bioactive surfaces, leading to ambiguous results of binding assays and negating the binding specificity of patterned cell-adhesive motives. Localized specific binding is often achieved by attaching a linker to the surface, and the other side of the linker is used to bind specifically to a desired functional agent, as e.g. proteins, antibodies, and fluorophores, depending on the function required by the application. We present a protein-repellent polymer brush enabling highly specific covalent surface immobilization of biorecognition elements by strain-promoted alkyne-azide cycloaddition click chemistry for selective protein adhesion. The protein-repellent polymer brush is functionalized by highly localized molecular binding sites in the low micrometer range using polymer pen lithography (PPL). Because of the massive parallelization of writing pens, the tunable PPL printed patterns can span over square centimeter areas. The selective binding of the protein streptavidin to these surface sites is demonstrated while the remaining polymer brush surface is resisting nonspecific adsorption without any prior blocking by bovine serum albumin (BSA). In contrast to the widely used BSA blocking, the reactive polymer brushes are able to significantly reduce nonspecific protein adsorption, which is the cause of biofouling. This was achieved for solutions of single proteins as well as complex biological fluids. The remarkable fouling resistance of the polymer brushes has the potential to improve the multiplexing capabilities of protein probes and therefore impact biomedical research and applications.
具有抗蛋白特性的反应性表面，可促进局部特异性结合，这在生物医学领域的应用中是非常理想的。非特异性黏附会影响生物活性表面的功能，导致结合分析的结果不确定，并否定图案化细胞黏附基序的结合特异性。通过将连接子附着到表面上，通常可以实现局部特异性结合，连接子的另一侧用于特异性结合所需功能试剂，例如蛋白质、抗体和荧光染料，具体取决于应用所需的功能。我们提出了一种抗蛋白聚合物刷，通过应变促进的炔基-叠氮化物环加成点击化学实现生物识别元件的高度特异性共价表面固定，用于选择性蛋白质黏附。通过聚合物笔式光刻（PPL）在低微米范围内使用高度局部化的分子结合位点对聚合物刷进行功能化。由于书写笔的大规模并行化，可调 PPL 打印图案可以跨越平方厘米区域。在没有牛血清白蛋白（BSA）预先阻断的情况下，表面位点对蛋白质链霉亲和素的选择性结合得到了证明，而其余聚合物刷表面则抵抗非特异性吸附。与广泛使用的 BSA 阻断相比，反应性聚合物刷能够显著减少非特异性蛋白质吸附，这是非生物污染的原因。这在单种蛋白质溶液以及复杂生物流体中都得到了证实。聚合物刷的显著抗污染能力有可能提高蛋白质探针的多重检测能力，从而对生物医学研究和应用产生影响。
