Method for detecting DNA tetrahedron drug loading system through capillary electrophoresis
A technology of capillary electrophoresis and drug-carrying system, which is applied in the field of capillary electrophoresis detection of DNA tetrahedron drug-carrying system, can solve the problem of inability to detect DNA tetrahedron and its drug-carrying system at the same time, and achieves expanded application, high repeatability, and operation. simple effect
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Embodiment 1
[0027] 1. Synthesis of DNA tetrahedron:
[0028] The purchased (Sangon (Shanghai, China)) DNA single strand was centrifuged in a centrifuge (8000r / min, 3min), and then an appropriate amount of TM buffer was added to make the DNA single strand concentration 100 μM. Mix the four single strands forming the DNA tetrahedron at equimolar concentrations in TM buffer to form a DNA single strand sample with a final concentration of 1 μM; heat the prepared sample in a water bath at 95°C for 10 minutes, then rapidly cool down to 4 ℃, and kept in a refrigerator at 4 ℃ for 3 hours, the DNA tetrahedral structure can be obtained.
[0029] 2. Mix 5 μL of DNA tetrahedron and 1 μL of doxorubicin evenly, and incubate together for 14 hours at room temperature in the dark, to obtain free DNA tetrahedron and DNA tetrahedron-doxorubicin complex.
[0030] 3. Capillary electrophoresis detection: use an uncoated quartz capillary with an effective length of 35cm (total length 60cm, inner diameter 75μm)...
Embodiment 2
[0032] According to the method of Example 1, the DNA tetrahedral structure was uniformly mixed with excess doxorubicin (concentration ratio: 1:150), and co-incubated at room temperature and protected from light for different times. It can be seen that with the prolongation of the incubation time, the peak (P2) intensity of the DNA tetrahedron-doxorubicin complex appearing in the electropherogram is increasing ( figure 2 ).
Embodiment 3
[0034] According to the method of Example 1, the DNA tetrahedron-doxorubicin complex (concentration ratio of 1:150) was placed in a solution of pH=5 or pH=7 (deionized water, adjusted pH value with 1M HCl), The drug release of the DNA tetrahedron-doxorubicin complex under different conditions was reflected by monitoring the change of the peak intensity of the free DNA tetrahedron at different times. It can be seen that the DNA tetrahedron drug delivery system releases drugs faster under acidic conditions ( image 3 ), and relatively good stability under neutral conditions ( Figure 4 ).
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