Journal: Journal of Nanobiotechnology

Article Title: Dendritic cell-targeted liposomes for cancer immunotherapy via inhibition of aryl hydrocarbon receptor

doi: 10.1186/s12951-025-03756-6

Figure Lengend Snippet: Identification of SP65 with affinity for DCs and synthesis of SP65-conjugated lipids. ( A ) Phage-display was used to select peptides that bound to the recombinant mouse CD11c (mCD11c) and DCs, but not to CD3 + T cells. After the first five rounds of biopanning, the phage titer eluted from mCD11c increased 2,200-fold, and after the subsequent two rounds, the titer from CD3 + T cell-unbound/DC-bound fractions increased 4,000-fold compared with the first round. Pfu, plaque-forming units. (B ) Binding activity and specificity between individual phage clones and mCD11c or bovine serum albumin were tested in 5th round by ELISA. A 450 , optical density at 450 nm. ( C ) Binding activities between the phage clone 65 (PC65) or control phage and bone marrow derived dendritic cells (BMDCs) were analyzed by flow cytometry. ( D ) Hydrogen bonds between SP65 and mCD11c demonstrated in Pymol. Light pink: αX. Slate Blue: β2. Green: SP65. Yellow: hydrogen bond (red arrow highlighted). ( E ) Synthesis scheme of SP65-DOPE; The sequence of synthetic peptide 65 (SP65) is PRSMPSRHKSQRGGGC. ( F ) MALDI-TOF mass spectrum of SP65-DOPE under positive mode; upper: found MS; lower: calculated MS. ( G ) Synthesis scheme of Ctrl-DOPE; Derived from amino acid residue 561–572 of BSA, the sequence of control peptide is KATEEQLKTVMEGGGC. ( H ) MALDI-TOF mass spectrum of Ctrl-DOPE under negative mode; upper: found MS; lower: calculated MS

Article Snippet: Briefly, recombinant mouse CD11c (mCD11c) protein (R&D Systems, Cat# 7987-AX) was coated onto 96-well plates and exposed to 2 × 10 11 phages from the Ph.D.−12 library (New England Biolabs) in PBS buffer at 4 °C for 1 h. After washing, bound phages were eluted with RIPA buffer, amplified, and titrated using the ER2738 Escherichia coli strain.

Techniques: Recombinant, Binding Assay, Activity Assay, Clone Assay, Enzyme-linked Immunosorbent Assay, Control, Derivative Assay, Flow Cytometry, Sequencing, Residue

Journal: Journal of Nanobiotechnology

Article Title: Dendritic cell-targeted liposomes for cancer immunotherapy via inhibition of aryl hydrocarbon receptor

doi: 10.1186/s12951-025-03756-6

Figure Lengend Snippet: SP65 enhances liposomal Dil uptake into DCs. ( A ) Fluorescence Dil uptake of splenic DCs after 30 min–4 h of incubation with variable formulations of liposomal Dil, n = 3. ( B ) Quantitative data of mean fluorescence intensity (MFI) of Dil in splenic DCs after 4 h of incubation with lipo-Dil (non-conjugated), Ctrl-lipo-Dil (control peptide-conjugated), or SP65-lipo-Dil (SP65-conjugated) from ( A ). ( C ) Confocal microscopic images of Dil uptake in splenic DCs. Cell shape was observed using the differential interference contrast model. (Scale bar = 50 μm). ( D ) In vivo Dil uptakes in CD11c + and CD11c − cells from peripheral blood of mice receiving liposomal Dil through intravenous injection. ( n = 3 biologically independent mice). ( F ) Quantitative data of the MFI of Dil in CD11c + cells from ( E ) after 2 h of injection. ( G ) Distribution of DiR in major organs from mice receiving liposomal DiR through intravenous injection after 2 h. ( G ) Quantitative data of ( F ) ( n = 4 biologically independent mice). All data were expressed as the mean values ± SD. Statistical differences were evaluated using one-way ANOVA. *, p < 0.05, **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. All statistical analyses were performed with GraphPad Prism 10.4.0

Article Snippet: Briefly, recombinant mouse CD11c (mCD11c) protein (R&D Systems, Cat# 7987-AX) was coated onto 96-well plates and exposed to 2 × 10 11 phages from the Ph.D.−12 library (New England Biolabs) in PBS buffer at 4 °C for 1 h. After washing, bound phages were eluted with RIPA buffer, amplified, and titrated using the ER2738 Escherichia coli strain.

Techniques: Fluorescence, Incubation, Control, In Vivo, Injection