Synthesis 2A1E-CBA-PEI

0.055 g 2-amino-1-ethanol

0.26 g CBA

2.5 ml 25% MeOH

60 oC for 2 days.

Then add 0.138 g PEI18 in 2 ml 25% MeOH, 45 oC for 24 hr.

Plate 3, 2A1E-CBA-PET18 vs PHP-DSP-PEI18

2A1E: 2-amino-1-ethanol

2A1E-CBA-PEI18, 1:10	2A1E-CBA-PEI18, 1:20	2A1E-CBA-PEI18, 1:30	2A1E-CBA-PEI18, 1:40
2A1E-CBA-PEI18, 1:50	2A1E-CBA-PEI18, 1:75	2A1E-CBA-PEI18, 1:100	2A1E-CBA-PEI18, 1:150
PHP-DSP-PEI18, 1:50	PHP-DSP-PEI18, 1:75	PHP-DSP-PEI12, 1:75	PHP-DSP-PEI18, 1:30, 11/10/2011

36 hours after transfection

4 days

Transfection efficiency is low.

only 1:10 has somewhat good expression

Protocol 2

PEI-CBA

Branched PEI2 (2k).

1. Dissolve 20 mg PEI2  in 5 ml MeOH/H2O solution (9:1, v/v) and stirr in a three-necked flask equipped with a condenser at 45◩C under N2 .

2. Add  different amount of CBA solution in methanol dropwise into polymer solutions.

3. Stir the mixture in the dark at 45◩ C under N2 atmosphere for 24 hr

3. Add  10% (w/w) excess PEI2 (1 mg/ml) to consume any un-reacted CBA, stir for another 6 h.

4. Dilute to 10 ml with distilled water and acidified to pH 4 with 10 M HCl.

5. Dialyse s (MWCO = 3500) against distilled water for 48 h.

6. Lyophilise for 24 h to give white powder.

PEI2–PA

N -acylation of PEI2K with PA  Palmitoyl chloride (PA)

1. Dissolve PEI2 (100 mg) in CHCl3 (100 ml) and stir in a three-necked flask under N2 .

2. After 30 min of stirrng, add triethylamine (80 ÎŒl) to the solution, stir for 30 min.

3. Cool the reaction mixture to approx. 4◩ C.

4. Add different amounts of PA solution (10, 25 and 50 ÎŒl of 1 mg/ml in CHCl3 ) drop-wise to reaction mix under stirring.

5. Stir at room temperature for 12 h.

6. Collect product by precipitation in excess ether.

7. Lyophilization.

--Paul.xu 14:45, 17 November 2011 (EST)