Protein surface patterning is employed in a broad spectrum of applications ranging from protein microarray analysis to 2D cell organization. However, limitations arise because of the highly sensitive nature of proteins requiring careful handling to ensure their structural and functional integrity during the grafting process. Here, we describe a patterning protocol that keeps proteins in an aqueous environment during their immobilization, avoiding the loss of their biological activity. The procedure is based on the UV-mediated removal of polyethylene glycol self-assembled monolayers in a transparent microfluidic chamber, giving access to micrometric motifs of predefined geometries. Afterward, modified proteins can be grafted on the photopatterned domains. We also studied the influence of reactive oxygen species for a better understanding of the chemical mechanism involved in this process. Finally, as a proof of concept, a protein microarray was created with this process using cell-capturing antibodies to immobilize human blood cells, confirming the functionality of the arrayed proteins.

中文翻译：

---

光学辅助表面功能化，用于蛋白质在水性介质中的排列

蛋白质表面图案化用于从蛋白质微阵列分析到2D细胞组织的广泛应用。但是，由于蛋白质的高度敏感性质而需要进行认真处理以确保其在嫁接过程中的结构和功能完整性，因此存在局限性。在这里，我们描述了一种模式化协议，该协议在固定化过程中将蛋白质保持在水性环境中，避免了其生物活性的损失。该程序基于在透明微流体腔室中紫外线介导的聚乙二醇自组装单层膜的去除，从而可以访问预定义几何形状的测微图案。之后，可以将修饰的蛋白质嫁接到光图案化的域上。我们还研究了活性氧的影响，以更好地了解此过程中涉及的化学机理。最终，作为概念验证，通过该过程使用捕获细胞的抗体固定人血细胞来创建蛋白质微阵列，从而证实了所排列蛋白质的功能。