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Preparation method of alginate/epsilon-polylysine/alginate biological microcapsule

An alginate gel, alginate technology, applied in microcapsule preparation, microsphere preparation, microcapsule and other directions, can solve the problem of limited application of sodium alginate/α-polylysine microcapsule, poor biocompatibility , high price and other problems, to achieve the effect of low cost, mild reaction conditions and safe use

Active Publication Date: 2013-09-18
ZHANGJIAGANG IND TECH RES INST CO LTD DALIAN INST OF CHEM PHYSICS CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the many biological microcapsules, sodium alginate / polylysine microcapsules are used more, and the microcapsule membrane material is chemically synthesized α-polylysine (the amide bond between lysine residues is made of α-amino and α-carboxyl condensation), but due to the high price of α-polylysine (300-400US$ / g), and the inherent poor biocompatibility of the material, it has certain toxicity, which greatly limits the use of algae. The clinical application of sodium lysate / α-polylysine microcapsules [Strand B L, Ryan TL, Veld P I, et al. Cell Transplant., 2001, 10: 263-275]

Method used

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  • Preparation method of alginate/epsilon-polylysine/alginate biological microcapsule
  • Preparation method of alginate/epsilon-polylysine/alginate biological microcapsule
  • Preparation method of alginate/epsilon-polylysine/alginate biological microcapsule

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Effect test

Embodiment 1

[0025] 1) Preparation by high-voltage electrostatic method [Qu Beibei, Yu Weiting, Liu Xiudong, etc., preparation of monodisperse calcium alginate microgel beads by high-voltage electrostatic method, Acta Chemical Industry, 2005, 56(8): 1547] for embedding Calcium alginate gel microspheres with CHO cells, the initial cell seeding density was 1×10 6 / mL sodium alginate solution.

[0026] 2) Under the condition of 37°C, immerse the calcium alginate gel microspheres prepared above into 0.05% (w / v), pH5.5, ε-polylysine (Mw10KDa) / normal saline solution, CHO / seaweed The volume ratio of calcium acid gel microspheres and ε-PLL solution was 1:40, reacted for 10 minutes to prepare ε-AP microcapsules, and then washed 3 times with normal saline.

[0027]3) Under the temperature condition of 37°C, immerse the ε-AP microcapsules prepared above in a sodium alginate (Mw200KDa) / 0.9% (w / v) NaCl solution with a concentration of 0.05% (w / v), CH0 / ε -AP microcapsules and sodium alginate solution ...

Embodiment 2

[0032] 1) Using high-voltage electrostatic method [Qu Beibei, Yu Weiting, Liu Xiudong, etc., preparation of monodisperse calcium alginate microgel beads by high-voltage electrostatic method, Acta Chemical Industry, 2005, 56(8): 1547] to prepare embedding Calcium alginate gel microspheres with hepatocytes-HepG2 cells, the initial cell seeding density was 1×10 6 / mL sodium alginate solution.

[0033] 2) Under the condition of 37°C, immerse the calcium alginate gel microspheres prepared above into 0.1% (w / v), pH6.5, ε-polylysine (Mw30KDa) / normal saline solution, HepG2 / seaweed The volume ratio of calcium acid gel microspheres and ε-PLL solution was 1:20, reacted for 15 minutes to prepare ε-AP microcapsules, and then washed 3 times with normal saline.

[0034] 3) Under the temperature condition of 37°C, immerse the ε-AP microcapsules prepared above in the sodium alginate (Mw300KDa) / 0.9% (w / v) NaCl solution with a concentration of 0.1% (w / v), HepG2 / ε The volume ratio of -AP microc...

Embodiment 3

[0038] 1) Using high-voltage electrostatic method [Qu Beibei, Yu Weiting, Liu Xiudong, etc., preparation of monodisperse calcium alginate microgel beads by high-voltage electrostatic method, Acta Chemical Industry, 2005, 56(8): 1547] to prepare embedding Calcium alginate gel microspheres with embryonic stem cells (ES) at an initial seeding density of 1×10 6 / mL sodium alginate solution.

[0039] 2) Under the condition of 37°C, immerse the calcium alginate gel microspheres prepared above into 0.2% (w / v), pH6.5, ε-polylysine (Mw50KDa) / normal saline solution, ES / seaweed The volume ratio of calcium acid gel microspheres and ε-PLL solution was 1:10, reacted for 15 minutes to prepare ε-AP microcapsules, and then washed 3 times with normal saline.

[0040] 3) Under the temperature condition of 37°C, immerse the ε-AP microcapsules prepared above in the sodium alginate (Mw400KDa) / 0.9% (w / v) NaCl solution with a concentration of 0.2% (w / v), ES / ε - AP microcapsules and sodium alginate ...

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Abstract

An alginate-e-polylysine microcapsule for embedding bioactive substances and its preparation method are disclosed. The microcapsule is a sphere with particle size of 100-1000 µm, which comprises microcapsule film and inner core. The microcapsule film is a polyelectrolyte complex hydrogel film obtained by reacting alginate withe-polylysine via static complexation. The inner core is a fluid- or hydrogel-environment comprising bioactive substances. The microcapsule is used as the carrier for cell transplantation, cell culture or embedding bioactive substances such as protein, polypeptide, enzyme or nucleic acid.

Description

technical field [0001] The invention relates to biological microcapsules, in particular to a preparation method of alginate / ε-polylysine / alginate biological microcapsules. Background technique [0002] In the 1960s, Chang reported semi-permeable membrane microcapsules, pointing out that biologically active substances and cells such as proteins and enzymes can be embedded in them to maintain the activity of biological substances [Chang TMS.Semipermeablemicrocapsules, Science, 1964, 146: 524-525] . In the early 1980s, Lim and Sun successfully prepared sodium alginate / α-polylysine (alginate / α-polylysine) semipermeable membrane microcapsules (referred to as α -APA microcapsules), encapsulating islet cells of Wistar rats and transplanted into diabetic WistarLewis rats to secrete and release insulin to regulate blood sugar [Lim F, Sun A M. Microencapsulated islets bioartificialendocrine pancreas, Science, 1980, 210: 908- 910]. This has promoted the rapid development of research...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J13/20A61K9/50A61K47/36A61K47/34C12N11/04
CPCA61K9/5036A61K9/5052B01J13/10A61K2035/128
Inventor 马小军朱静于炜婷张英
Owner ZHANGJIAGANG IND TECH RES INST CO LTD DALIAN INST OF CHEM PHYSICS CHINESE ACADEMY OF SCI
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