Fig. S1.
Characterization of anti-CD47 nanobodies isolated from an alpaca-derived phage VHH library. (A) Sequences of four nanobody clones isolated from the phage library panned against mouse CD47 ECD. (B) SDS/PAGE analysis of recombinant anti-CD47 nanobody clones isolated from E. coli and purified via affinity and size exclusion chromatography. nr, nonreducing; r, reducing. (C) FACS plots demonstrating 100 nM mouse SIRPa tetramers (Alexa-647–labeled) binding to LSTRA cell surface CD47 in the absence (Top Right) or presence of 1 μM anti-CD47 nanobody clones (Bottom). The gate shown is drawn based on background staining with negative control Alexa-647 tetramers (Top Left).