Cellular therapies remain constrained by the limited availability of sensors for disease markers. Here we present an integrated target-to-receptor pipeline for constructing a customizable advanced modular bispecific extracellular receptor (AMBER) that combines our generalized extracellular molecule sensor (GEMS) system with a high-throughput platform for generating designed ankyrin repeat proteins (DARPins). For proof of concept, we chose human fibrin degradation products (FDPs) as markers with high clinical relevance and screened a DARPin library for FDP binders. We built AMBERs equipped with 19 different DARPins selected from 160 hits, and found 4 of them to be functional as heterodimers with a known single-chain variable fragments binder. Tandem receptors consisting of combinations of the validated DARPins are also functional. We demonstrate applications of these AMBER receptors in vitro and in vivo by constructing designer cell lines that detect pathological concentrations of FDPs and respond with the production of a reporter and a therapeutic anti-thrombotic protein.

细胞疗法仍然受到用于疾病标志物的传感器的有限可用性的限制。在这里，我们提出了一个集成的目标到受体管道，用于构建可定制的高级模块化双特异性细胞外受体 (AMBER)，该管道将我们的通用细胞外分子传感器 (GEMS) 系统与用于生成设计锚蛋白重复蛋白 (DARPins) 的高通量平台相结合。为了证明概念，我们选择人纤维蛋白降解产物 (FDP) 作为具有高度临床相关性的标记物，并筛选了 DARPin 文库中的 FDP 结合剂。我们构建了配备 19 种不同 DARPins 的 AMBER，从 160 次命中中选择，并发现其中 4 种具有已知单链可变片段结合剂的异二聚体功能。由经过验证的 DARPins 组合组成的串联受体也具有功能。