Fig. 2: Preparation, optimization, and anti-protein-adsorption effect of AFMFlips.

a, b Preparation and optimization of AFMFlips with serum-resistance membrane-fusion capacity. Liposomes were labeled with 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindodicarbocyanine,4-chlorobenzenesulfonate salt (DiD). The nuclei were stained with DAPI and the cell membranes were labeled with CellMask. The experiment was repeated three times independently with similar results. c Composite comparison of AFMFlip and the previously reported MFlips (MFlip-a and MFlip-b). d, e Hydrodynamic sizes (d) and zeta potentials (e) of AFMFlip, MFlip-a, and MFlip-b after 1-h incubation in the medium without or with 10% FBS at 37 °C. Data are presented as mean ± SD and statistically analyzed using two-tailed t test (n = 3 biologically independent samples). ns: no significance (p > 0.05). Measurements were conducted in water at room temperature. f Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of corona proteins on AFMFlip, MFlip-a, and MFlip-b after 1-h incubation in the medium with 10% FBS or 10% mouse serum. FBS, fetal bovine serum; 1-F, AFMFlip incubated with 10% FBS; 2-F, MFlip-a incubated with 10% FBS; 3-F, MFlip-b incubated with 10% FBS; MS, mouse serum; 1-M, AFMFlip incubated with 10% mouse serum; 2-M, MFlip-a incubated with 10% mouse serum; 3-M, MFlip-b incubated with 10% mouse serum. The experiment was repeated three times independently with similar results. g The bicinchoninic acid (BCA) quantification of protein corona from AFMFlip, MFlip-a, and MFlip-b after 1-h FBS incubation with different concentrations. Data are presented as mean ± SD (n = 3 biologically independent samples). h The area under the curve (AUC) analysis based on the BCA-quantification results from g. Data are presented as mean ± SD and statistically analyzed using one-way ANOVA (n = 3 biologically independent samples). i–l Protein classification for corona proteins recovered from AFMFlip, MFlip-a, and MFlip-b after 1-h incubation in 10% FBS-containing medium by liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. The identified proteins from corona were classified based on isoelectric point (pI) (i), molecular weight (MW) (j), gravy index (k), and instability index (l). Source Data are provided as a Source Data file.