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Pharmaceutical Care and Research: 2018; 18(2):87-91,95
DOI: 10.5428/pcar20180202
Construction and in vitro evaluation of A33 antibody functionalized exosomes drug delivery system
1. LI Yan1(1.School of Pharmacy,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China
2. 2(2.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China
3. GAO Shen2( 2.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China )
4. XIA QingMing2( 2.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China )
5. GONG ChunAi2( 2.Department of Pharmacy,Changhai Hospital,Second Military Medical University,Shanghai 200433,China )
6. GUO HuiLing1(1.School of Pharmacy,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China
ABSTRACT  Objective: To prepare and evaluate in vitro A33 antibody functionalized exosomes(EXO) loading doxorubicin(DOX) for drug delivering.Methods:A33 antibodies were coupled to carboxyl-ultrasmall superparamagnetic iron oxide nanoparticles(USPIO) by chemical crosslinking method to prepare A33-USPIO.Size distribution and zeta potential of USPIO and A33-USPIO were determined by dynamical light scattering,and their structures were elucidated by infrared spectroscopy(IR).EXO from human colon carcinoma cell line LIM1215 cells were isolated by ultracentrifugation and the morphology was observed by transmission electron microscope after DOX loading.Enzyme linked immunosorbent assay(ELISA) was used to detect the optimum proportion of A33-USPIO and EXO.Drug release rate of A33-US-EXO/DOX was examined by dialysis method.Cellular uptake was investigated by flow cytometry after different DOX formulations were co-cultured with LIM1215 cells.Fluorescence microscopy was used to observed the fluorescence in A33 antigen positive and negative cells to verify the targeting ability of A33-US-EXO/DOX.Results:Under transmission electron microscope,the diameter of EXO was about 100 nm,and 32 μg EXO could bind to 2.5 μg A33-USPIO.After binding,the particle size and polymer dispersity index(PDI) of A33-US-EXO/DOX were (187.83±6.76) nm and 0.21,respectively.At the conditions of pH 7.4,6.0 and 5.0,the cumulative releases of DOX from A33-US-EXO/DOX in 48 h were 24.32%,34.07% and 62.82%,respectively.The uptake efficiency of cells treated with DOX,EXO/DOX and A33-US-EXO/DOX were (13.10±1.08)%,(51.53±3.56)% and (85.11±3.91)%,respectively.The fluorescence images showed that a large amount of A33-US-EXO/DOX bound with A33 antigen positive LIM1215 cells,while a very small amount bound with A33 antigen nagetive RAW264.7 cells.Conclusion:The A33 antigen targeting drug delivery system was successfully constructed and showed a high uptake efficiency and good targeting ability in vitro.
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LI Yan1,2,GAO Shen2,XIA QingMing2,GONG ChunAi2,GUO HuiLing1,. Construction and in vitro evaluation of A33 antibody functionalized exosomes drug delivery system[J]. Pharmaceutical Care and Research / yao xue fu wu yu yan jiu. 2018; 18(2): 87-91,95.
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