Paul.xu at 20:01, 17 November 2011

2011-11-17T20:01:25Z

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

Diethylenetriamine, abbreviated dien or DETA, (H2N-CH2CH2-NH-CH2CH2-NH2, an analog of diethylene glycol) Triethylenetetramine, abbreviated trien or TETA, (H2N-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH2) Tetraethylenepentamine, abbreviated TEPA, (H2N-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH-CH2CH2-NH2), 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. 

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

 

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

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

 

 

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

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

 

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

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

 

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

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

 

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

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

 

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

 

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

 

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

 

[[Image:Transfection efficiency SS-PBAA-PEI.jpg|600px|Transfection efficiency SS-PBAA-PEI.jpg]]

 

 

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

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

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

3, Purify by ultrafiltration (MWCO 1,000) 

4, Freeze dry 

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

6, Transfection: 

Dissolve in 20 mM HEPES buffered saline 

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

Serum free medium for 3~4 hr 

then add complete serum

TEPA-CBA has the best transfection efficiency

 

'''Reducible TATA by Michaelâs-type addition''' 

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. 

 

1, Add  CBA (0.8 g, 3.07 mmol) to TETA (449 mg, 3.07 mmol) in 10% methanol (2.8 mL) 

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

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

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 

[[Image:Tetraethylenepentamine.jpg]] 

Tetraethylenepentamine (TEPA) 

 

[[Image:Ethylenediamine.jpg]] 

Ethylenediamine (EDA) 

 

[[Image:Triethylenetetramine.jpg]] 

Triethylenetetramine (TETA) 

 

[[Image:Diethylenetriamine.jpg]] 

Diethylenetriamine (DETA) 

[[Image:Reducible.jpg]]

[[Image:Amido amines.jpg]]

[[Image:Polymers.jpg]]

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

Synthesis of TEPA CBA - Revision history

Paul.xu at 20:01, 17 November 2011