Supplementary date: Application of Gyrolab microfluidic platform to measure picomolar affinity of a PD-L1-binding Adnectin PET radioligand

Supplementary Figure 1. Bioanalytical characterization of Adnectin samples. (A) SDS-PAGE: lane 1, standard; lane 3, His-ADX_5322_A02-DBCO; lane 5, ADX_5322_A02-DBCO; lane 6, 19F-BMS-986192; lanes 2 and 4, other constructs. (B) Analytical SEC. (C) Mass spectrometry (MS). Supplementary Figure 2. Multi-curve analysis of the affinity series for PD-L1 Adnectins after 22 h or 44 h equilibration at room temperature (data points and fitted curve: 50 pM Adnectin in orange and 150 pM Adnectin in blue). (A) ADX_5322_A02-DBCO, 44 h; (B) His-ADX_5322_A02-DBCO, 22 h; (C) His-ADX_5322_A02-DBCO, 44h; (D) 19F-BMS-986192, 22 h; (E) 19F-BMS-986192, 44 h.

补充图1. 阿德奈汀（Adnectin）样品的生物分析表征。(A) 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）：泳道1为标准品；泳道3为His-ADX_5322_A02-DBCO；泳道5为ADX_5322_A02-DBCO；泳道6为19F-BMS-986192；泳道2和泳道4为其他重组构建体。(B) 分析型尺寸排阻色谱（Analytical SEC）。(C) 质谱（Mass spectrometry, MS）。补充图2. 室温下平衡22小时或44小时后，PD-L1靶向阿德奈汀亲和力梯度的多曲线分析（数据点与拟合曲线：50 pM阿德奈汀以橙色标注，150 pM阿德奈汀以蓝色标注）。(A) ADX_5322_A02-DBCO，平衡44小时；(B) His-ADX_5322_A02-DBCO，平衡22小时；(C) His-ADX_5322_A02-DBCO，平衡44小时；(D) 19F-BMS-986192，平衡22小时；(E) 19F-BMS-986192，平衡44小时。