Antibody synthetic bacteria-nano stimulant heterozygote system and application thereof
A stimulant, nano-technology, applied in the application field of cytokine nano-stimulator, antibody synthetic bacteria-nano-stimulator hybrid system, can solve problems affecting the function and activity of living cells, and achieve immune-related adverse reactions Severe, addressing effects of low immune response rates
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Embodiment 1
[0094] Embodiment 1: Mal-poly(PEG 77 -co-His) preparation
[0095] Michael addition reaction to prepare tumor extracellular pH-responsive copolymers, such as figure 2 shown. Weigh a certain amount of 1,6-hexanediol diacrylate (HDDA), histamine (His) and maleimide-polyethylene glycol amino (Mal-PEG 77 -NH 2 ) were mixed in a molar ratio of 1:0.85:0.15 and dissolved in a certain volume of dimethyl sulfoxide (DMSO), reacted for 48 hours at 55°C in an argon atmosphere, purified and freeze-dried to obtain a polymer powder. The copolymer is further analyzed and identified by gel permeation chromatography / NMR.
Embodiment 2
[0096] Embodiment 2: DBCO-poly(PEG 77 -co-His) preparation
[0097] Michael addition reaction to prepare tumor extracellular pH-responsive copolymers, such as image 3 shown. Weigh a certain amount of 1,6-hexanediol diacrylate (HDDA), histamine (His) and diphenylcyclooctyne-polyethylene glycol amino (DBCO-PEG 77 -NH 2 ) were mixed in a molar ratio of 1:0.85:0.15 and dissolved in a certain volume of dimethyl sulfoxide (DMSO), reacted for 48 hours at 55°C in an argon atmosphere, purified and freeze-dried to obtain a polymer powder. The copolymer is further analyzed and identified by gel permeation chromatography / NMR.
Embodiment 3
[0098] Example 3: Preparation of Mal-labeled IL-12 nano-stimulator
[0099] Preparation of IL-12 nano-stimulator by microemulsion method: 10 μg of IL-12 was dissolved in a small amount of ultrapure water, 10 mg of Mal-poly(PEG 77 -co-His) copolymer was dissolved in 10 times the equivalent volume of dichloromethane, mixed and stirred for 1min, added 1% polyvinyl alcohol (PVA) and stirred for 1min, and finally added 10mL PBS and continued to stir for 30min to obtain IL-12 nano-stimulator agent solutions (INPs). The surface Zeta potential, particle size and PDI were detected by DLS, the morphology was observed by TEM and SEM, and the drug loading and encapsulation efficiency were detected by IL-12p70ELISA kit.
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