Editing Synthesis of CBA-PolyAmines (section)

== Synthesis<br>  ==

Dissolve SP (0.202g, 1 mmol) and Dde-OH (0.419 g, 2.3 mmol)&nbsp; in 1 mL MeOH, stirring at room temperature for 24 h to let Dde group specifically protect primary amine groups in spermine.<br> 

The next day, add CBA (0.260g, 1 mmol) into above system in 1.5 mL of MeOH/diH 2 O (9/1 v/v). <br> 

Polymerize at 60° C in the dark under nitrogen atmosphere for 3-4 days.<br> 

Add 10% mol excess EDA&nbsp; to consume any unreacted acrylamide groups, stirring at 60°C for at least additional 2 h. <br> 

Precipitate polymer in 40 mL anhydrous diethyl ether to get the intermediate polymer poly(CBA-SP-Dde). <br> 

Remove the Dde groups in the deprotection mixture of NH2OH·HCl/Imidazole/NMP/DMF by stirring at room temperature for 4 h. <br> 

'''To make deprotection mixture: '''<br> 

1.25 g (1.80 mmol) of NH2OH·HCl <br> 

0.918 g (1.35 mmol) of imidazole is suspended in 5 mL of NMP. <br> 

Sonicate the mixture until complete dissolution. <br> 

Just before reaction, dilute 5 volume of this solution&nbsp; with 1 volume of DMF to make the deprotection mixture. <br> 

After deprotection, purify the crude product by dialysis (MWCO = 1000) against MilliQ water for 24 h<br> 

Dry by lypholization to obtain poly(CBA-SP) as solid gel.<br> 

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[[Image:Polyamine CBA synthesis.png|600px|Polyamine CBA synthesis.png]]<br> 

[[Image:Polyamine CBA cytotoxicity.png|600px|Polyamine CBA cytotoxicity.png]]<br> 

[[Image:Polyamine CBA TR efficiency.png]]<br> 

poly(CBA-TETA) and poly(CBA-AEPD)is the best for uptake<br> 

but poly(CBA-SP), poly(CBA-APPD) and poly(CBA-APED) have higher transfection efficiency<br> 

for cost + efficiency: use poly(CBA-APED)<br> 

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