Fig. 1

Design, preparation and characterization of NanoBE. (a) Schematic illustration of recombinant lentivirus LV-𝛂CD40/CLDN18.2 scFv plasmid. The scFv sequence was fused to the C-terminus of the signal peptide and the N-terminus of PDGFR transmembrane domain. The cells were transfected with the constructed recombinant lentivirus. (b) Flow cytometry analysis (left) and confocal imaging (right) of mCherry-positive cells showed the 𝛂CD40 scFv and 𝛂CLDN18.2 scFv expression. (c) Preparation of Nano/CD40, Nano/CLDN18.2 and NanoBE. Nano/CD40 and Nano/CLDN18.2 was prepared by coating cell membrane from KPC-𝛂CD40 scFv and KPC-𝛂CLDN18.2 scFv onto PLGA nanoparticle core, respectively. NanoBE was prepared using cell membrane derived from both KPC-𝛂CD40 scFv and KPC- 𝛂CLDN18.2 scFv. (d) TEM images of PLGA alone and NanoBE. Scale bar = 100 nm. (e) SDS-PAGE protein analysis of PLGA, cell membrane isolated from KPC-𝛂CD40/𝛂CLDN18.2 scFv, Nano/CD40, Nano/CLDN18.2, and NanoBE. (f) Flow cytometry analysis of the mCherry signal on Nano/CLDN18.2 and Nano/CD40. (g) Flow cytometry analysis of NanoBE containing both DiD-labeled KPC-𝛂CD40 scFv and DiO-KPC-𝛂CLDN18.2 scFv membrane. Gray, the particles without cell membrane; pink, the particles with both cell membrane. (h) Particle size and (i) 𝛇-potentials of PLGA alone and NanoBE