A kind of anti-glioma polypeptide molecule and its application
A technology for glioma and glioma cells, which is applied in the direction of polypeptides containing localization/targeting motifs, anti-tumor drugs, peptide/protein components, etc., and can solve problems such as inability to accurately target gliomas
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Embodiment 1
[0027] The purity of TAT-PICK1 was analyzed by analytical high performance liquid chromatography (flow rate: 1ml / min). Specifically:
[0028] The model of the analytical high-performance liquid chromatograph: Agilent 1200, the model of the chromatographic column used: AngilentEclipse XDB-C18 Analytical, 5um, 4.6X150006Dm.
[0029] Chromatographic operating conditions: linear gradient elution, the eluent is composed of mobile phase A liquid and B liquid. Mobile phase A liquid: 0.1% by volume concentration of trifluoroacetic acid aqueous solution (water as solvent), mobile phase B liquid: 0.1% by volume concentration of trifluoroacetic acid in acetonitrile solution (acetonitrile as solvent). The elution time was 12 minutes in total, the elution flow rate was 1 mL / min, and the ultraviolet detection wavelength was 220 nm.
[0030] The detection result of analytical high performance liquid chromatography showed that the purity of the obtained TAT-PICK1 was 97.2%.
Embodiment 2
[0032] The chemical structure of TAT-PICK1 was characterized by MALDI-TOF mass spectrometry, and the mass spectrometry characterization results of TAT-PICK1 were as follows figure 2 shown. Molecular weight (Mw): 2825.30, the other three peaks can also prove that the molecular weight of TAT-PICK1 is consistent.
Embodiment 3
[0034] TAT-PICK1 polypeptide molecule inhibits the migration of glioma cell C6
[0035] Transwell plate (Corning Company, product number #3422) was used to detect the effect of TAT-PICK1 polypeptide on the C6 migration ability of glioma cells. Cell migration experiments were performed according to the instructions of the Transwell plate. The main operation steps are as follows:
[0036] 1) Dilute the digested and counted glioma cell C6 and TAT-PICK1 polypeptide to 200,000 / mL with serum-free culture medium DMEM (final concentration is 30uM), take 200uL and add it to the upper chamber of the Transwell, and add 600uL to the lower chamber of the Transwell to completely The cell culture solution was used as the experimental group; the serum-free culture solution containing the TAT leader peptide was used as the negative control group.
[0037] 2) in CO 2 37°C, 5% CO in an incubator 2 Continue to culture for 24 hours under the same conditions, scrape off the cells in the upper T...
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