Engineered by nature, biological entities are exceptional building blocks for biomaterials. These entities can impart enhanced functionalities on the final material that are otherwise unattainable. However, preserving the bioactive functionalities of these building blocks during the material fabrication process remains a challenge. We describe a high-throughput protocol for the bottom-up self-assembly of highly concentrated phages into microgels while preserving and amplifying their inherent antimicrobial activity and biofunctionality. Each microgel is comprised of half a million cross-linked phages as the sole structural component, self-organized in aligned bundles. We discuss common pitfalls in the preparation procedure and describe optimization processes to ensure the preservation of the biofunctionality of the phage building blocks. This protocol enables the production of an antimicrobial spray containing the manufactured phage microgels, loaded with potent virulent phages that effectively reduced high loads of multidrug-resistant Escherichia coli O157:H7 on red meat and fresh produce. Compared with other microgel preparation methods, our protocol is particularly well suited to biological materials because it is free of organic solvents and heat. Bench-scale preparation of base materials, namely microporous films (the template for casting microgels) and pure concentrated phage suspension, requires 3.5 h and 5 d, respectively. A single production run, that yields over 1,750,000 microgels, ranges from 2 h to 2 d depending on the rate of cross-linking chemistry. We expect that this platform will address bottlenecks associated with shelf-stability, preservation and delivery of phage for antimicrobial applications, expanding the use of phage for prevention and control of bacterial infections and contaminants.

生物实体是自然设计的，是生物材料的特殊构建模块。这些实体可以赋予最终材料增强的功能，这是其他方式无法实现的。然而，在材料制造过程中保留这些构件的生物活性功能仍然是一个挑战。我们描述了一种高通量方案，用于将高度浓缩的噬菌体自下而上自组装成微凝胶，同时保留和放大其固有的抗菌活性和生物功能。每个微凝胶由五十万个交联的噬菌体作为唯一的结构成分组成，自组织成对齐的束。我们讨论了制备过程中的常见缺陷，并描述了确保噬菌体构建模块的生物功能得以保留的优化过程。该方案能够生产含有制造的噬菌体微凝胶的抗菌喷雾，该喷雾含有强毒力噬菌体，可有效减少红肉和新鲜农产品上的多重耐药大肠杆菌O157:H7 的高含量。与其他微凝胶制备方法相比，我们的方案特别适合生物材料，因为它不含有机溶剂和热量。基础材料的小规模制备，即微孔膜（铸造微凝胶的模板）和纯浓缩噬菌体悬浮液，分别需要 3.5 小时和 5 d。单次生产运行可产生超过 1,750,000 个微凝胶，所需时间从 2 小时到 2 天不等，具体取决于化学交联速率。我们预计该平台将解决与抗菌应用噬菌体的货架稳定性、保存和交付相关的瓶颈，扩大噬菌体在预防和控制细菌感染和污染物方面的用途。