Editing Synthesis of TEPA CBA

[[Image:SS-PAEI synthesis.jpg]] 

Diethylenetriamine, abbreviated dien or DETA, (H2N-CH2CH2-NH-CH2CH2-NH2, an analog of diethylene glycol)<br> Triethylenetetramine, abbreviated trien or TETA, (H2N-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH2)<br> Tetraethylenepentamine, abbreviated TEPA, (H2N-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH2),<br> Pentaethylenehexamine, abbreviated PEHA, (H2N-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH2), up to polyethylene amine. Similarly piperazine is an analogue of dioxane.<br><br>

[[Image:Chemical structure of bioreduciable polymers.jpg]] <br> [[Image:Linear paa vs branched paa.jpg|650px|Linear paa vs branched paa.jpg]]<br> 

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[[Image:Hyper branched CBA-EDA.jpg|650px|Hyper branched CBA-EDA.jpg]]<br> <br> 

[[Image:Reducible hyperbranched poly amido amine)2.jpg|650px|Reducible hyperbranched poly amido amine)2.jpg]]<br> 

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[[Image:Poly(amindoamine)s structure.jpg]]<br> 

[[Image:CBA-DAH-API co polymer.jpg|650px|CBA-DAH-API co polymer.jpg]]<br> 

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[[Image:EDA-CBA-DNA complex.jpg|650px|EDA-CBA-DNA complex.jpg]]<br> 

[[Image:TETA-CBA-VEGF-Suppression.jpg|650px|TETA-CBA-VEGF-Suppression.jpg]]<br> 

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[[Image:Serum interaction DNA protection.jpg|650px|Serum interaction DNA protection.jpg]]<br> 

[[Image:Transfection efficiency-EDA-CBA.jpg|650px|Transfection efficiency-EDA-CBA.jpg]]<br> 

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[[Image:Reducible cationic polymers example.jpg|650px|Reducible cationic polymers example.jpg]] 

[[Image:Hyper branched polymers.jpg|650px|Hyper branched polymers.jpg]] 

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[[Image:Ss-pAA.jpg|650px|Ss-pAA.jpg]] 

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[[Image:Instable DSP.jpg|650px|Instable DSP.jpg]] 

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[[Image:CBA-DMEDA-polymer.jpg|650px|CBA-DMEDA-polymer.jpg]] 

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'''Syntheis of Reducible TEPA-CBA copolymer''' 

1. Tetraethylenepentamine (2.2 mmol) +&nbsp; bisacrylate CBA (0.58 mg, 2.2 mmol) &nbsp; in 2.2 ml 10&nbsp;% aqueous methanol at 45 °C for 3~4 days. <br> 

2, Add excess amount of TEPA, 20 mol% excess and react for additional 3 days<br> 

3, Purify by ultrafiltration (MWCO 1,000)<br> 

4, Freeze dry<br> 

5, Analysis: 1H-NMR&nbsp; and gel permeation chromatography (GPC), MW:3,350. 

6, Transfection:<br> 

Dissolve in 20 mM HEPES buffered saline<br> 

'''1.5:1''' (polymer:DNA) in PBS or serum free medium, complex: 15~30 min 

Serum free medium for 3~4 hr<br> 

then add complete serum 

TEPA-CBA has the best transfection efficiency 

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'''Reducible TATA by Michaelâs-type addition'''<br> 

Reaction: ethylene amine monomer (TETA, DETA, or EDA) and CBA in 10% aqueous MeOH at 50 °C in the dark for a period of 18 h to 2 days under an argon atmosphere. <br> 

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1, Add&nbsp; CBA (0.8 g, 3.07 mmol) to TETA (449 mg, 3.07 mmol) in 10% methanol (2.8 mL)<br> 

2, The polymerization ((50 °C) until stirring of the magnetic bar becomes difficult due to the high viscosity of the reaction mixture (16 h).<br> 

3, Add excess TETA (âŒ50 mg) in MeOH to consume any remaining acrylamide functional groups, stir for an additional 2 h.<br> 

4, Add 10 ml ultrapure deionized water (ddH2O), adjust pH to 5, isolate polymer product by ultrafiltration (MWCO 1000). 

5, Freeze dry 

6, DNA:polymer=1:3, incubate 15~30 min 

7, Serum-free medium for 4 hours, then replace with complete medium<br><br> 

[[Image:Tetraethylenepentamine.jpg]]<br> 

Tetraethylenepentamine (TEPA)<br> 

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[[Image:Ethylenediamine.jpg]]<br> 

Ethylenediamine (EDA)<br> 

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[[Image:Triethylenetetramine.jpg]]<br> 

Triethylenetetramine (TETA)<br> 

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[[Image:Diethylenetriamine.jpg]]<br> 

Diethylenetriamine (DETA)<br> 

[[Image:Reducible.jpg]] 

[[Image:Amido amines.jpg]] 

[[Image:Polymers.jpg]] 

[[Category:Transfection]] [[Category:Paul]]