Drug-loaded nanoparticles for targeted immunotherapy of pancreatic cancer and its application
A nanoparticle and pancreatic cancer technology, applied in gene therapy, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problems of limited in vivo application and lack of effective carriers
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Embodiment 1
[0022] The targeting carrier of this example includes CD44v6 single chain antibody, and nanocarrier IONP-PEI, which carries siBmi-1. The content of this embodiment includes two parts: experimental method and conclusion analysis.
[0023] 1. Experimental method
[0024] 1. Cell culture
[0025] Human pancreatic cancer cell line Panc-1 was cultured in high-glucose DMEM medium containing 10% fetal bovine serum at a constant temperature of 37°C and 5% CO 2 cultured in an incubator.
[0026] 2. Targeting carrier construction
[0027] scFvCD44v6 (CD44v6 single-chain antibody) was synthesized according to the methods disclosed in Chinese patents 2011102402903 and 2011102402871. ScFvCD44v6 is coupled to PEI at a mass ratio of 1:1, and the process is briefly described as follows:
[0028] ① Prepare the IONP-PEI complex according to the ratio, dissolve mal-PEG-NHS in sterilized deionized water, add scFv to the complex at a molar ratio of 1:10, mix well, and incubate at room tempera...
Embodiment 2
[0176] The targeting carrier of this example includes CD44v6 single-chain antibody and nanocarrier IONP-PEI, which co-load gemcitabine and siBmi-1. The content of this embodiment includes two parts: experimental method and conclusion analysis.
[0177] 1. Experimental method
[0178] 1. Cell culture
[0179] Human pancreatic cancer cell line Panc-1 was cultured in high-glucose DMEM medium containing 10% fetal bovine serum at a constant temperature of 37°C and 5% CO 2 cultured in an incubator.
[0180] 2. Preparation and characterization of Gem-IONP (carrier IONP carrying gemcitabine) and svFv-Gem-IONP (targeting carrier carrying gemcitabine and siBmi-1)
[0181] (1) In an ice bath, add triethylamine (Triethylamine) and acid-sensitive tetrapeptide (9–fluoronylmethoxycarbonyl(Fmoc)-Gly-Phe-Leu-Gly(GFLG)-nitrophenyl ester peptide) according to 7.5umol: 6.8μmol To 1.0mol acetonitrile solution.
[0182] (2) Gemcitabine and triethylamine (Triethylamine) were dissolved in 1 ml o...
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