Dendritic human-derived cell-penetrating peptide hpp7k, its production and its method for mediating plasmid dna transfection
A membrane-penetrating peptide, human-derived technology, applied in the field of biomedicine, can solve the problems of insecurity and low transfection efficiency, and achieve the effects of low cost, low immunogenicity, and easy quality control
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Embodiment 1
[0048](1) Weigh a certain amount of Fmoc-Arg(PBF)-Wang resin or H-Arg(PBF)-2cl resin, pour it into a glass reactor, add an appropriate amount of dichloromethane (DCM) to soak until the resin swells, and pump Dry;
[0049] (2) Add an appropriate amount of deprotection solution to the reactor, agitate with nitrogen for 0.5-2 hours, drain, add an appropriate amount of dimethylformamide (DMF) to the reactor, agitate with nitrogen, drain, repeat operation 1-10 Second-rate;
[0050] (3) Weigh the amino acid in each step equivalent to 1 times the molar amount of the resin, 1 times the molar amount of the carbodisubtype condensing agent (such as dicyclohexylcarbodiimide (DCC)) or benzotriazolium salt Type condensing agent (such as O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) or O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboron Hydrochloric acid (TBTU)) was dissolved in dimethylformamide (DMF) or DCM or tetrahydrofuran (THF) and added to the reactor, reac...
Embodiment 2
[0058] (1) Weigh a certain amount of Fmoc-Arg(PBF)-Wang resin or H-Arg(PBF)-2cl resin, pour it into a glass reactor, add an appropriate amount of dichloromethane (DCM) to soak until the resin swells, and pump Dry;
[0059] (2) Add an appropriate amount of deprotection solution to the reactor, agitate with nitrogen for 0.5-2 hours, drain, add an appropriate amount of DMF into the reactor, agitate with nitrogen, drain, and repeat the operation 1-10 times;
[0060] (3) Weigh amino acids equivalent to 10 times the molar amount of the resin in each step, 10 times the molar amount of the carbon disubtype condensing agent (such as dicyclohexylcarbodiimide (DCC)) or benzotriazolium salt Type condensing agent (such as O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) or O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboron Hydrochloric acid (TBTU)) was dissolved in dimethylformamide (DMF) or DCM or tetrahydrofuran (THF) and added to the reactor, reacted for 1-4h, a...
Embodiment 3
[0068] (1) Weigh a certain amount of Fmoc-Arg(PBF)-Wang resin or H-Arg(PBF)-2cl resin, pour it into a glass reactor, add an appropriate amount of dichloromethane (DCM) to soak until the resin swells, and pump Dry;
[0069] (2) Add an appropriate amount of deprotection solution to the reactor, agitate with nitrogen for 0.5-2 hours, drain, add an appropriate amount of DMF into the reactor, agitate with nitrogen, drain, and repeat the operation 1-10 times;
[0070] (3) Weigh the amino acid that is equivalent to 5 times the molar amount of the resin, the carbodisubtype condensing agent (such as dicyclohexylcarbodiimide (DCC)) or the benzotriazolium salt of 5 times the molar amount Type condensing agent (such as O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) or O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboron Hydrochloric acid (TBTU)) was dissolved in dimethylformamide (DMF) or DCM or tetrahydrofuran (THF) and added to the reactor, reacted for 1-4h, and...
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