Cationized polysaccharide nanoparticle gene delivery systems and manufacturing method thereof
A gene delivery system and cationization technology, applied in the field of cationized polysaccharide nanoparticle gene delivery system, can solve the problem of not being directly used as a gene, without positive charge, etc., achieving no immunogenicity, good encapsulation and release effect, wide The effect of the application foreground
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Embodiment 1
[0055] Example 1: Preparation of the gene delivery system of the cationized Pleurotus eryngii polysaccharide-DNA plasmid nanocomposite modified by spermine
[0056] Weigh 0.5g refined Pleurotus eryngii polysaccharide, dissolve in 30ml double distilled water, add 0.75g KIO 4 , quickly placed in a dark room, stirred magnetically, and reacted at room temperature for 72 hours; adding 10ml ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the above conditions; put the reaction solution into a dialysis bag (cutoff molecular weight > 3500Da), and dialyzed in double distilled water for 48 hours ; The dialysate was freeze-dried to obtain the oxidized Pleurotus eryngii polysaccharide.
[0057] Take 0.15g of the above-mentioned oxidized Pleurotus eryngii polysaccharide and dissolve it in 10ml of double distilled water; weigh 0.3g of spermine and dissolve it in 5ml of borate buffer solution (pH=9); use a disposable...
Embodiment 2
[0059] Example 2: Preparation of a gene delivery system of ethylenediamine-modified cationized Pleurotus eryngii polysaccharide-DNA plasmid nanocomposite
[0060] Weigh 0.6 g refined Pleurotus eryngii polysaccharide, dissolve in 50ml double distilled water, add 0.89g KIO 4 , quickly placed in a dark room, magnetically stirred, and reacted at room temperature for 72 hours; adding 12ml of ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the above conditions; put the reaction solution into a dialysis bag and dialyzed in double distilled water for 48 hours (molecular weight cut-off > 3500Da) ; The dialysate was freeze-dried to obtain the oxidized Pleurotus eryngii polysaccharide.
[0061] Take 0.4 g of oxidized Pleurotus eryngii polysaccharide and dissolve it in 30ml of double distilled water; take 0.7 ml of ethylenediamine and dissolve it in 5ml of borate buffer solution (pH=9); use a disposable syringe t...
Embodiment 3
[0063] Example 3: Preparation of the gene delivery system of cationized mulberry leaf polysaccharide-DNA plasmid nanocomposite modified by ethylenediamine
[0064] Weigh 0.8g refined mulberry leaf polysaccharide, dissolve in 50ml double distilled water, add 1.2g KIO 4 , quickly placed in a dark room, magnetically stirred, and reacted at room temperature for 72 hours; adding 12ml of ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the above conditions; put the reaction solution into a dialysis bag and dialyzed in double distilled water for 48 hours (molecular weight cut-off > 3500Da) ; The dialysate was freeze-dried to obtain the oxidized mulberry leaf polysaccharide.
[0065] Take 0.5g of oxidized mulberry leaf polysaccharide and dissolve it in 30ml of double distilled water; take 0.28ml of ethylenediamine and dissolve it in 5ml of borate buffer solution (pH=9); Add it into the mulberry leaf polysacch...
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