Therapeutic delivery · 2026
Target-specific daunorubicin nanoplatform based on anti-CD22-ScFv decorated β-cyclodextrin metal organic frameworks.
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Khabiri Alireza, Rastegari Banafsheh, Rafiei Dehbidi Gholamreza, Savardashtaki Amir, Takhshid Mohammad Ali
Аннотация
AIMS: This study aimed to develop a targeted drug delivery system based on anti-CD22 single-chain variable fragment (ScFv) conjugated to β-cyclodextrin (β-CD) Bio-MOFs for the selective delivery of daunorubicin (DNR) to CD22-positive B-cell tumor cells.
MATERIALS AND METHODS: The anti-CD22-ScFv gene was designed using the variable heavy (VH) and light (VL) chain regions of the human RFB4 antibody and cloned into the pET28a vector, followed by expression in E. coli BL21 (DE3). The recombinant protein was purified by affinity chromatography and confirmed by SDS-PAGE. Anti-CD22-ScFv-β-CD Bio-MOFs were synthesized using glutaraldehyde and L-lysine as crosslinkers and subsequently loaded with DNR. The formation of the nanostructure was verified by ATR-FTIR spectroscopy and zeta potential analysis. Drug uptake and cytotoxic effects were evaluated in CD22-positive Raji cells and CD22-negative Jurkat cells using fluorescence microscopy, flow cytometry, and the MTT assay.
RESULTS: Successful expression and purification of anti-CD22-ScFv were confirmed by SDS-PAGE. Characterization analyses verified the formation of anti-CD22-ScFv-β-CD-DNR Bio-MOFs. The targeted system facilitated efficient uptake of DNR in CD22-positive Raji cells and induced significant dose-dependent cytotoxicity. In contrast, minimal effects were observed in CD22-negative Jurkat cells.
CONCLUSIONS: Anti-CD22-ScFv-β-CD-DNR Bio-MOFs demonstrate effective and selective delivery of DNR to CD22-expressing cells and may represent a promising targeted therapeutic approach for B-cell malignancies.
MATERIALS AND METHODS: The anti-CD22-ScFv gene was designed using the variable heavy (VH) and light (VL) chain regions of the human RFB4 antibody and cloned into the pET28a vector, followed by expression in E. coli BL21 (DE3). The recombinant protein was purified by affinity chromatography and confirmed by SDS-PAGE. Anti-CD22-ScFv-β-CD Bio-MOFs were synthesized using glutaraldehyde and L-lysine as crosslinkers and subsequently loaded with DNR. The formation of the nanostructure was verified by ATR-FTIR spectroscopy and zeta potential analysis. Drug uptake and cytotoxic effects were evaluated in CD22-positive Raji cells and CD22-negative Jurkat cells using fluorescence microscopy, flow cytometry, and the MTT assay.
RESULTS: Successful expression and purification of anti-CD22-ScFv were confirmed by SDS-PAGE. Characterization analyses verified the formation of anti-CD22-ScFv-β-CD-DNR Bio-MOFs. The targeted system facilitated efficient uptake of DNR in CD22-positive Raji cells and induced significant dose-dependent cytotoxicity. In contrast, minimal effects were observed in CD22-negative Jurkat cells.
CONCLUSIONS: Anti-CD22-ScFv-β-CD-DNR Bio-MOFs demonstrate effective and selective delivery of DNR to CD22-expressing cells and may represent a promising targeted therapeutic approach for B-cell malignancies.