Peptides are promising drug candidates because of their diversity, biocompatibility and spectrum of activities. Here, we describe a protocol for high-throughput screening of SPOT-peptide arrays to assess the antibiofilm, antimicrobial and immunomodulatory activities of synthetic peptides. It is a Protocol Extension of our previous Nature Protocols article, which describes the synthesis of SPOT-peptide arrays and assays for screening antimicrobial activity. This latest protocol allows the simultaneous assessment of hundreds of synthetic host defense peptides to define their overall activity profiles and identify candidate sequences that are suitable for further characterization and development as anti-infectives. When coupled with the SPOT-array technology for peptide synthesis, the described procedures are rapid, inexpensive and straightforward for peptide library screening. The protocols can be implemented in most microbiology or immunology research laboratories without the need for specialists. The time to complete each step ranges between 1 and 4 h with overnight pauses, and datasets related to the antibiofilm and immunomodulatory activities of a large set of peptide sequences can be generated in a few days.

肽因其多样性、生物相容性和活性谱而成为有前途的候选药物。在这里，我们描述了一种用于高通量筛选 SPOT 肽阵列的协议，以评估合成肽的抗生物膜、抗菌和免疫调节活性。它是我们之前自然协议的协议扩展文章，描述了 SPOT 肽阵列的合成和用于筛选抗菌活性的测定。这一最新协议允许同时评估数百种合成宿主防御肽，以定义它们的整体活性谱并确定适合进一步表征和开发作为抗感染剂的候选序列。当与用于肽合成的 SPOT 阵列技术相结合时，所描述的程序对于肽库筛选来说是快速、廉价和直接的。该协议可以在大多数微生物学或免疫学研究实验室中实施，而无需专家。完成每个步骤的时间介于 1 到 4 小时之间，并有隔夜停顿，