Liposome complexes consisting of TM4SF5 B cell epitope and CpG-DNA (MB-ODN 4531(O)) co-encapsulated with DOPE: CHEMS were prepared as reported previously (12) (link). Briefly, DOPE and CHEMS were mixed in 10% ethanol at a molar ratio of 1：1, evaporated with nitrogen gas to make a solvent-free lipid film, and resuspended in a mixture containing equal volumes of water-soluble MB-ODN 4531(O) (50 μg) and peptide (50 μg), followed by vigorous stirring at room temperature for 30 min. After adjusting the pH to 7.0, the complex of peptide and Lipoplex(O) was sonicated lightly for 30 s with a sonicator (Sonifier 450, Branson Ultrasonics). After the complex was filtered with a 0.22 μm filter, it was freeze-thawed three times with liquid nitrogen.
Protocol detail
Co-encapsulated Liposome Complexes for Immunotherapy
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Corresponding Organization :
Other organizations : Hallym University, Chungbuk National University, Yonsei University
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Variable analysis
independent variables
- Molar ratio of DOPE and CHEMS (1:1)
- Concentration of MB-ODN 4531(O) (50 μg)
- Concentration of peptide (50 μg)
- Characteristics of the liposome complexes (not explicitly mentioned)
- Solvent (10% ethanol)
- Mixing and stirring conditions (room temperature for 30 min)
- PH adjustment (to 7.0)
- Sonication (30 s)
- Filtration (0.22 μm)
- Freeze-thaw cycles (three times)
- Positive control: Not mentioned
- Negative control: Not mentioned
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