Currently, high-throughput approaches are lacking in the isolation of antibodies with functional readouts beyond simple binding. This situation has impeded the next generation of cancer immunotherapeutics, such as bispecific T cell engager (BiTE) antibodies or agonist antibodies against costimulatory receptors, from reaching their full potential. Here, we developed a highly efficient droplet-based microfluidic platform combining a lentivirus transduction system that enables functional screening of millions of antibodies to identify potential hits with desired functionalities. To showcase the capacity of this system, functional antibodies for CD40 agonism with low frequency (<0.02%) were identified with two rounds of screening. Furthermore, the versatility of the system was demonstrated by combining an anti-Her2 × anti-CD3 BiTE antibody library with functional screening, which enabled efficient identification of active anti-Her2 × anti-CD3 BiTE antibodies. The platform could revolutionize next-generation cancer immunotherapy drug development and advance medical research.

目前，在分离具有超越简单结合的功能读数的抗体方面缺乏高通量方法。这种情况阻碍了下一代癌症免疫疗法，例如双特异性 T 细胞接合剂 (BiTE) 抗体或针对共刺激受体的激动剂抗体，无法充分发挥其潜力。在这里，我们开发了一种高效的基于液滴的微流体平台，结合了慢病毒转导系统，可以对数百万种抗体进行功能筛选，以识别具有所需功能的潜在命中。为了展示该系统的能力，通过两轮筛选鉴定了具有低频率 (<0.02%) 的 CD40 激动作用的功能性抗体。此外，该系统的多功能性通过将抗 Her2 × 抗 CD3 BiTE 抗体库与功能筛选相结合来证明，从而能够有效识别活性抗 Her2 × 抗 CD3 BiTE 抗体。该平台可以彻底改变下一代癌症免疫治疗药物的开发并推进医学研究。