Paul.xu at 19:01, 13 November 2011

2011-11-13T19:01:08Z

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[[Image:Paa_synthesis.jpg]]

[[Image:EDA-CBA_synthesis.jpg]]

Synthesis of Poly(amido amine)s (PAAs) homopolymers by Michael addition

'''p(HMBA-AEP) and p(CBA-AEP)'''

1. Add HMBA(0.55 g, 2.46 mmol) or CBA (0.64 g, 2.46 mmol) and AEP (0.32 g, 2.46 mmol) into a brown reaction flask with methanol as a solvent (2.5 mL).

2. Let polymerization in the dark at 40°C under nitrogen atmosphere.

3. The reaction mixture will become homogeneous in less than 1 hour, allow the reaction for 6 days. You will see a viscous solution.

4. Add 10 mol% excess AEP (0.03 g, 0.2 mmol) to consume any unreacted acrylamide groups, let stir for 2 days at 40°C.

5, Dilute the resulting solution with water to about 30 ml, acidify with 6 M HCl to pH~4, and then dialyze through a dialysis membrane (MWCO 1000g/mol) for 3 days.

6. Freeze-drying. p(MBA-AEP) and p(BAP-AEP)

Polymerization in water as the solvent, terminate the reactions in 6 hours and 3 hours, respectively.

[[Image:CBA APOL synthesis.jpg]]

 

 

[[Image:CBA AEP synthesis.jpg]]

co-polymer

'''[[Image:CBA AEP synthesis2.jpg]]'''

mono-polymer

 '''[[Image:PABOL structure.jpg]] p(HMBA80/CBA20-AEP) Copolymers''': PAA copolymers with varying amounts of disulfide groups

AEP + HMBA and CBA at various ratios (i.e. 80/20, 60/40 and 40/60).

1. Add CBA (0.16 g, 0.60 mmol), HMBA (0.54 g, 2.41 mmol) and AEP (0.39 g, 3.01 mmol) into a brown reaction flask with methanol as a solvent (3.0 mL).

2. Stir the reaction mixture at 40°C in the dark under nitrogen atmosphere for 6 days.

3. Add 10 mol% excess of AEP (0.04 g, 0.03 mmol) to consume any unreacted acrylamide groups, stir for 2 days at 40°C.

4, Dilute the resulting solution with water to about 30 ml.

5, Acidify with 6 M HCl to pH~4.

6. Dialyze through dialysis membrane (MWCO 1000g/mol) for 3 days.

7. Freeze-drying the copolymer to get HCl-salt.

p(CBA-AEP) has best transfection efficiency, 1:3 to 1:6. CBA: 100% has the best transfection efficiency.

'''Bioreducible poly(amido amine)s (SS-PAAs).'''

 1. Add N, Nâ-cystaminebisacrylamide CBA (0.250 g, 1.1 mM) and 5-amino-1-pentanol (APOL) (1.1 mM) to a brown reaction flask.

2. Add methanol/water mixture (1.1 mL, 4/1, v/v).

3. Polymerize in the dark at 45 °C under nitrogen atmosphere for 10 days yielding a viscous solution.

4. Add 10 mol% excess 5-amino-1-pentanol to consume any unreacted acrylamide, stir for 2 days at 45 °C.

5. Dilute with 30 ml water, acidify with 6 M HCl aqueous solution to pH~3.

6. Purify by ultrafiltration with acidic deionised water (pH~4).

7, freeze-dry to yield solid powder.

[[Image:Transfection efficiency SS-PAA.jpg]]

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Synthesis of PAA - Revision history

Paul.xu at 19:01, 13 November 2011