微生物病原体，包括细菌、真菌和病毒，可以对临床使用的药物产生耐药性；因此，寻找新的治疗药物是一个持续的挑战。最近，我们报道了光免疫抗菌策略（PIAS），这是一种光免疫技术，能够从分子角度靶向消除多种微生物，包括病毒病原体严重急性呼吸综合征冠状病毒2和多重耐药细菌病原体耐甲氧西林金黄色葡萄球菌 (MRSA)。 PIAS 的工作原理与光免疫疗法 (PIT) 相同，自 2020 年以来，光免疫疗法已在日本用于治疗复发性头颈癌。PIAS 和 PIT 均使用与酞菁衍生物染料缀合的单克隆抗体，该染料在光激活时会发生形状变化。这种形状变化会引起抗体-染料缀合物的结构变化，从而导致缀合物的结合位点内产生物理应力并破坏它们。因此，靶向准确性和灵活性可以根据所用抗体的特异性来确定。在本协议中，我们描述了如何设计治疗策略、用染料标记单克隆抗体并表征产品。我们提供了如何在体外和体内设置和执行 PIAS 和 PIT 应用程序的详细示例。这些例子包括以MRSA为代表的针对微生物的PIAS、在VeroE6/TMPRSS2细胞中使用严重急性呼吸综合征冠状病毒2针对病毒的PIAS、针对MRSA感染动物的PIAS以及针对癌细胞的体外和体内PIT。体外和体内方案可分别在约 3 小时和 2 周内完成。© 2023。Springer Nature Limited。

Microbial pathogens, including bacteria, fungi and viruses, can develop resistance to clinically used drugs; therefore, finding new therapeutic agents is an ongoing challenge. Recently, we reported the photoimmuno-antimicrobial strategy (PIAS), a type of photoimmunotechnology, that enables molecularly targeted elimination of a wide range of microbes, including the viral pathogen severe acute respiratory syndrome coronavirus 2 and the multidrug-resistant bacterial pathogen methicillin-resistant Staphylococcus aureus (MRSA). PIAS works in the same way as photoimmunotherapy (PIT), which has been used to treat recurrent head and neck cancer in Japan since 2020. Both PIAS and PIT use a monoclonal antibody conjugated to a phthalocyanine derivative dye that undergoes a shape change when photoactivated. This shape change induces a structural change in the antibody-dye conjugate, resulting in physical stress within the binding sites of the conjugate and disrupting them. Therefore, targeting accuracy and flexibility can be determined based on the specificity of the antibody used. In this protocol, we describe how to design a treatment strategy, label monoclonal antibodies with the dye and characterize the products. We provide detailed examples of how to set up and perform PIAS and PIT applications in vitro and in vivo. These examples are PIAS against microbes using MRSA as a representative subject, PIAS against viruses using severe acute respiratory syndrome coronavirus 2 in VeroE6/TMPRSS2 cells, PIAS against MRSA-infected animals, and in vitro and in vivo PIT against cancer cells. The in vitro and in vivo protocols can be completed in ~3 h and 2 weeks, respectively.© 2023. Springer Nature Limited.