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).

<b>Supplementary
Figure 2. </b>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. 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，标准品；泳道3，His-ADX_5322_A02-DBCO；泳道5，ADX_5322_A02-DBCO；泳道6，¹⁹F-BMS-986192；泳道2和4，其他构造体。(B)分析型尺寸排阻色谱（analytical SEC）。(C)质谱分析（mass spectrometry, MS）。

<b>补充图2：</b>PD-L1 Adnectin亲和力系列的多曲线分析，在室温下平衡22小时或44小时（数据点与拟合曲线：50 pM Adnectin为橙色，150 pM Adnectin为蓝色）。(A) ADX_5322_A02-DBCO，44小时；(B) His-ADX_5322_A02-DBCO，22小时；(C) His-ADX_5322_A02-DBCO，44小时；(D) ¹⁹F-BMS-986192，22小时；(E) ¹⁹F-BMS-986192，44小时。