Preparation method of phospholipid biomimic membrane containing aquaporins
An aquaporin and biomimetic membrane technology, applied in chemical instruments and methods, membrane technology, semi-permeable membrane separation, etc., can solve the problem of not being able to improve water flux and retention rate at the same time, and achieve the effect of broad application prospects.
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Embodiment 1
[0024] Mix 2 ml of 1,2-dioleoylphosphatidylcholine at a concentration of 1 mg / ml with 2 ml of a chloroform solution of 1,2-dioleyloxy-3-trimethylaminopropane at a concentration of 1 mg / ml, and wash with Blow dry with nitrogen and vacuum dry, then add 30 ml of PBS buffer solution with pH=7.5, shake and sonicate to prepare mixed phospholipid vesicle solution of two phospholipids; add aquaporin 1 (0.008mg / ml / ) / glycerol (0.1mg / ml) / n-octyl-β-D-glucopyranoside (0.01mg / ml) PBS buffer solution 5ml, put the mixed solution into a dialysis bag with a molecular weight cut-off of 7KDa, and then buffer in PBS Dialyze in the solution for 3 days to prepare proteoliposomes with a ratio of protein to phospholipid of 1:50; immerse the polyacrylonitrile membrane prepared by the phase inversion method in 1.5M NaOH solution for 1.5h, wash it with deionized water, and remove the membrane Alternately soak in polyethyleneimine solution and sodium polystyrene sulfonate solution to obtain a composite me...
Embodiment 2
[0026] Mix 2 ml of 1,2-dioleoylphosphatidylcholine at a concentration of 1 mg / ml with 2 ml of a chloroform solution of 1,2-dioleyloxy-3-trimethylaminopropane at a concentration of 1 mg / ml, and wash with Blow dry with nitrogen and vacuum dry, then add 30 ml of PBS buffer solution with pH=7.5, shake and sonicate to prepare the mixed phospholipid vesicle solution of the above two phospholipids; add aquaporin 1 (0.008mg / ml / ) / Glycerol (0.1mg / ml) / n-octyl-β-D-glucopyranoside (0.01mg / ml) PBS buffer solution 2.5ml, put the mixed solution into a dialysis bag with a molecular weight cut-off of 7KDa, and then in Dialyze in PBS buffer solution for 3 days to prepare proteoliposomes with a ratio of protein to phospholipid of 1:100; immerse the polyacrylonitrile membrane prepared by the phase inversion method in 1.5M NaOH solution for 1.5h, wash it with deionized water, Soak the membrane alternately in polyethyleneimine solution and sodium polystyrene sulfonate solution to obtain a layer of ...
Embodiment 3
[0028]Mix 2 ml of 1,2-dioleoylphosphatidylcholine at a concentration of 1 mg / ml with 2 ml of a chloroform solution of 1,2-dioleyloxy-3-trimethylaminopropane at a concentration of 1 mg / ml, and use Blow dry with nitrogen and vacuum dry, then add 30 ml of PBS buffer solution with pH=7.5, shake and sonicate to prepare the mixed phospholipid vesicle solution of the above two phospholipids; add aquaporin 1 (0.008mg / ml / ) / Glycerol (0.1mg / ml) / n-octyl-β-D-glucopyranoside (0.01mg / ml) PBS buffer solution 1.25ml, put the mixed solution into a dialysis bag with a molecular weight cut-off of 7KDa, and then in Dialyze in PBS buffer solution for 3 days to prepare proteoliposomes with a ratio of protein to phospholipid of 1:200; immerse the polyacrylonitrile membrane prepared by the phase inversion method in 1.5M NaOH solution for 1.5h, wash it with deionized water, Soak the membrane alternately in polyethyleneimine solution and sodium polystyrene sulfonate solution to obtain a layer of assemb...
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