CN114134065B - 生物胞膜***及其制备方法、应用 - Google Patents
生物胞膜***及其制备方法、应用 Download PDFInfo
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- CN114134065B CN114134065B CN202111164850.1A CN202111164850A CN114134065B CN 114134065 B CN114134065 B CN 114134065B CN 202111164850 A CN202111164850 A CN 202111164850A CN 114134065 B CN114134065 B CN 114134065B
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Abstract
本发明公开一种生物胞膜***及其制备方法、应用。生物胞膜***包括磷脂双分子层、多胺复合物和聚异戊二烯类化合物。多胺复合物包括游离或聚合状态下的多胺复合物。多胺复合物选自如下化合物的双端分别具有两个氨基的直链或支链烷基胺、双端分别具有至少两个氨基且包含1‑4个仲胺的直链或支链烷基胺、一端具有至少一个氨基另一端为咪唑基的直链或支链烷基胺、环状多胺中的任意一种或多种。聚异戊二烯类化合物选自四萜化合物、四萜化合物脂肪酸酯衍生物中的任意一种或多种。本发明提供的生物胞膜***,能够有效降低糖脂含量,且能获得多胺复合物和聚异戊二烯类化合物等有利成分的含量较高,从而提升生物适应性,应用范围更广。
Description
技术领域
本发明涉及生物和高分子材料技术领域,尤其涉及一种生物胞膜***及其制备方法、应用。
背景技术
生物膜或生物性膜是细胞、细胞器和其环境接界的所有膜结构的总称,起着划分和分隔细胞和细胞器的作用,也是与细胞内通讯有关的重要部位。早年为便于研究,往往采用单一或几种脂质组成的人工膜。目前,虽然人工膜已在实际中得到广泛应用,但是人工膜存在着以下缺点:在体外易氧化、渗漏、储存性不好;在体内易被一些酶类物质降解和巨噬细胞吞噬而不能到达靶组织发挥有效作用等缺点,这些都限制了其作为载体的应用。此外,人工合成物质的添加,使得人工膜作为高分子材料植入人体不可避免的会使机体产生排异,使用受限。
天然生物膜虽然可以有效避免人工膜的缺陷,但是,由于天然生物膜的成分复杂,离纯化难度较大,某些不利成分例如糖脂等物质会降低生物适应性,而某些有利成分例如类胡萝卜素、生物多胺、SOD等有利于生物修复。带正电荷多胺结构能与带负电的细胞膜表面相互吸附形成电偶,使得经这类生物膜包裹的有效成分,能够更有效地经过内吞作用被导入细胞内而发挥功效作用。但是,现有技术难以同时做到在保证不利成分含量较低的前提下,能够不降低甚至提高有利成分的含量。
发明内容
本发明的目的在于提供一种能够有效降低糖脂含量,且能获得多胺复合物和聚异戊二烯类化合物等有利成分的含量较高的生物胞膜***,从而提升生物适应性,应用范围更广。
本发明采取的技术方案如下:本发明提供一种生物胞膜***,包括磷脂双分子层以及接枝于所述磷脂双分子层的多胺复合物和镶嵌于所述磷脂双分子层上的聚异戊二烯类化合物。其中,所述多胺复合物包括任意一种或多种:双端分别具有两个氨基的直链或支链烷基胺,双端分别具有至少两个氨基且包含1-4个仲胺的直链或支链烷基胺,一端具有至少一个氨基另一端为咪唑基的直链或支链烷基胺,环状多胺。所述聚异戊二烯类化合物选自四萜化合物、四萜化合物脂肪酸酯衍生物中的任意一种或多种的组合。
其中,四萜化合物包括但不限于类胡萝卜素,主要可以是嗜热玉米黄素、嗜热叶黄素脂肪酸酯、β-隐黄质花生酸酯。
磷脂双分子层与皮肤的细胞膜结构类似,能够起到肌肤调理的功能。多胺复合物具有促进抗辐射、抗氧化、促进细胞生长等多种功能。聚异戊二烯类化合物具有抗辐射、抗氧化等多种功能。此外,生物胞膜***中的多胺复合物带正荷,提高了与带负电相关的细胞膜的亲和性,更有利于作为脂质体的包裹用途,同时也可用于细胞转运的用途。
本发明所述的磷脂双分子层包括但不限于:双层磷脂分子结构、嵌入的膜蛋白、通道和膜相连的细胞器。
长链的多胺类似蛋白***,也能够***磷脂双分子层上,但由于多胺复合物可与脂肪酸或蛋白的羧基结合,也可与磷脂的磷酸键结合,因此多胺复合物在磷脂酸分子层上主要以接枝的方式为主。类胡萝卜素(聚异戊二烯类化合物)则是镶嵌式***为主。
可选地,所述生物胞膜***还具有糖脂,所述糖脂的含量≤100ppm。糖脂的含量较低。
可选地,所述生物胞膜***还包括具有生物活性的氧化还原酶,所述氧化还原酶的活性中心具有金属离子;所述金属离子来自Mn、Ca、Mg、Zn、Cu、Fe中的任意一种或多种。其中,Mn、Ca、Mg、Zn、Cu、Fe离子选自如下金属元素的常见价态离子:Mn、Ca、Mg、Zn、Cu、Fe。所述氧化还原酶选自超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、琥珀酸脱氢酶(SDH)中的任意一种或多种。具体地,所述氧化还原酶为超氧化物歧化酶,且其活性中心具有锰离子。SOD具有抗氧化性能。其中,Mn-SOD热稳定性较为优异。
可选地,多胺复合物的主要形式为酰化聚合多胺,所述多胺复合物具体可以为组胺、胍丁胺、戊二胺、精脒、腐胺、亚精胺、去甲精胺、高精胺、精胺中的一种或多种的组合。多胺复合物包括游离状态或聚合状态。其中,去甲精胺为N-氨基丙基去甲精胺,亚精胺为N-氨基丙基-亚精胺。环状多胺为含卟啉结构的细胞色素c。其中,细胞色素选自细菌色素,具体可以为菌脂素。
本发明还提供一种上述的生物胞膜***的制备方法,具体包括以下步骤:
1)通过温度梯度法、调节光照强度、调节渗透压中的任一种或多种组合的方法在无糖培养基中进行菌株的高代谢培养;
2)收集菌体,采用匀浆缓冲液重悬菌体,离心,取沉淀的菌体进行破碎,得到细胞裂解液;
3)加入重悬介质中得到重悬液,通过超速离心法及二次密度梯度离心法对分层的重悬液中生物胞膜***进行提纯,得到所需的生物胞膜***。
细胞裂解液加入到不同浓度的重悬介质中,然后以150,000-300,000×g,60-90min,1-6℃条件下超速离心,收集所需区域的液体C。将液体C加PBS缓冲溶液稀释,以100,000-200,000×g,30-90min,1-6℃条件下超速离心,弃上清,收集沉淀,即为所需提取的生物胞膜***。
当重悬介质为蔗糖时,可以选择物质的量浓度分别为0.3-0.9mol/L、1-3mol/L和0.01-0.3mol/L的蔗糖溶液。其中,由于不同浓度的蔗糖溶液的密度不同,在离心时分步加入不同浓度的蔗糖溶液形成密度梯度。
除上述制备方法之外,本发明的生物胞膜***还可以通过下述制备方法,具体包括以下步骤:
1)通过温度梯度法、调节光照强度、调节渗透压中的任一种或多种组合的方法在无糖培养基中进行菌株的高代谢培养;
2)收集菌体,采用匀浆缓冲液重悬菌体,离心,取沉淀的菌体进行破碎,得到细胞裂解液;
3)在细胞裂解液中加入有机溶剂提取脂质,将提取的脂质干燥成薄膜,再将薄膜加水混匀后进行离心,弃沉淀,收集上清液A;
4)对上清液A进行超速离心,弃上清液,收集沉淀B,得到所需的生物胞膜***。
其中,有机溶剂为氯仿-甲醇溶液,有机溶剂可以提取脂溶性成分,从而降低糖脂等水溶性成分的含量。在生物胞膜***的提取时,先将提取的脂质干燥成薄膜,再将薄膜加水混匀后进行离心,干燥的目的是可以通过干燥除去有机溶剂。所述重悬介质由蔗糖、甘油、聚蔗糖、氯化铯、氯化钠、三碘苯甲酰葡萄糖胺、硅溶胶、氟代碳中的至少一种配制而成。
其中,菌株为嗜热栖热菌、嗜热链球菌、水生栖热菌、嗜酸热硫化菌、硫化叶菌、热变形菌、餐古菌、暖球形菌、除硫球菌、酵母菌、枯草芽孢杆菌中的任一种或其组合。
本申请中的菌株采用无糖、高渗透压、光照等严苛的培养条件,使得在培养过程中加速菌株的糖脂代谢能力。严苛的培养条件指的是在菌株能存活的前提下,提高菌株的代谢。
本发明所述的无糖培养基中的碳源为不含多羟基醛或多羟基酮及其缩聚物和某些衍生物等糖类成分,即不含各种糖、淀粉、糊精、糖醇,碳源选择可以是乳制品或蛋白质或少量纤维素或酵母膏等。
可选地,上述两种制备方法中的步骤1)通过添加渗透压调节溶质调节发酵液的渗透压至350-3100mOsmol/kg,所述渗透压调节溶质为盐或甘油。盐可以为NaCl或CaCl2。具体地,渗透压调节溶质为1%-10%(w/v)的NaCl溶液或1%-15%(w/v)的CaCl2溶液,或5-30%(w/v)的甘油溶液(甘油和去离子水配制而成)。在培养基的渗透压较高的情况下,可以加速糖脂的代谢。
可选地,上述两种制备方法中的步骤1)温度梯度法采用60-90℃的温度范围内采用逐步升温的方式培养24h。
可选地,上述两种制备方法中的步骤1)光照强度的调节范围为500-1500lx,光照时间为24-72h。嗜热栖热菌为不需要光照的生长菌,为提高菌的光照生长适应过程,采用梯度加强光照法或较低照度的恒光照射法,促使细菌调整生长代谢以适应新的环境。
上述两种方法制备得到的生物胞膜***中的各物质成分含量如下表1所示。
表1生物胞膜***的成分
物质 | 含量 |
膜相对密度 | 1.160-1.290 |
蛋白质(%) | 5-15 |
磷脂(mg/g) | 40-200 |
总多胺(mg/g) | 20-95(质量百分比) |
类胡萝卜素(ppm) | 50-500 |
RNA(%) | 0.03-1.0 |
SOD(U/g) | 20-400 |
糖脂(ppm) | 0.5-95 |
本发明提供的上述的生物胞膜***主要通过膜内包裹、表面吸附、表面交联、膜间嵌镶、靶向中的一种或者多种结合的方式对活性物质进行结合。
本申请的生物胞膜***的磷脂双分子层构成球状或者囊泡状的脂质体,具有包裹功能和靶向功能,可以作为载色体、细胞器囊泡等载体用途,也可以作为天然脂质体用途;还具有生物酶功能,能够部分运载生物成分(主动运转和被动转运)能力。
具体地,所述活性物质可以为疫苗、免疫调节剂、化妆品成分、药用成分、基因物质、细胞或细胞组织。
本发明的有益效果是:本发明提供了一种生物胞膜***的制备方法,通过对菌株采用光照、高渗透压的苛刻条件下采用无糖培养基进行培养,使得菌株在繁殖过程中尽可能地消耗糖脂,并且产生多胺复合物、类胡萝卜素、SOD等有利成分。同时,在生物胞膜提取过程中,由于糖脂为水溶性物质,而多胺复合物、类胡萝卜素、SOD等为脂溶性物质,通过有机溶剂提取或者二次密度梯度离心的方法,能够有效地减少提取的生物胞膜***中的糖脂的含量,且使得多胺复合物、类胡萝卜素、SOD等有利成分的含量较高。
附图说明
图1是本发明实施例3、实施例5和实施例6得到的不同生物胞膜***中的蛋白质电泳结果图;
图2是本发明实施例10中不同生物胞膜***对HaCaT细胞的光损伤修复对比图;
图3是本发明实施例11中不同浓度的生物胞膜***在LPS模型的抗炎能力对比图。
具体实施方式
下面结合各附图,通过具体实施例,对本发明进行详细、完整的描述。
本申请中,SOD的酶活力测定按GB5009.171的方法进行,以抑制邻苯三酚自氧化速率达50%时所对应的酶量为一个SOD活力单位(U)来计算。
总多胺的含量检测采用酰氯反应法(以多胺(PAS)在碱性条件,与苯甲酰氯反应,将苯甲酰基团连在一级和二级胺上),在波长254nm处测定。总多胺的含量包括了组胺、胍丁胺、戊二胺、精脒、腐胺、亚精胺、去甲精胺、高精胺、精胺和菌脂素的所有的多胺复合物的含量总和。
类胡萝卜素的含量检测采用GB/T5009.83的方法测定。
糖脂的含量以3-去氧-D-甘露-2-辛酮糖酸(Kdo)计,用硫代巴比妥酸法测定可得。
金属离子的含量检测采用原子吸收光谱仪(AAS)进行元素分析。
本申请实施例采用的菌种均为嗜热栖热菌,是从美国黄石国家公园温泉池中分离纯化而来,保藏编号为ATCC27634。嗜热栖热菌能够在高渗透压的苛刻环境中生存。将嗜热栖热菌按1:100的比例接种到无糖培养基中。
实施例1采用的菌种为嗜热栖热菌。
1)菌株培养:将嗜热栖热菌按1:100的比例接种到培养基中,利用梯度高热发酵法与高压、高渗透结合的方法,提高了多胺复合物、类胡萝卜素、SOD的产率。加NaCl至浓度为1.35%,调节发酵液渗透压至460mOsmol/kg,采用60-70℃,70-75℃,75-90℃每8H逐级升温,在终条件下,继续培养24h。
2)破壁:发酵后在4,000rpm,30min,4℃条件下离心收集菌体;用匀浆缓冲液(20mmol/L Tris-Cl pH8.0,100mmol/L NaCl,2mmol/L MgCl2,1mmol/L DTT)重悬菌体,在6,000rpm,10min,4℃条件下离心去上清;加入匀浆缓冲液重悬沉淀(约1g加入10ml缓冲液),再加入终浓度为1mmol/L的PMSF,冰浴超声破碎30min(振幅为55%,超声5s,停8s),得到细胞裂解液。
3)生物胞膜***提取:取10ml细胞裂解液用氯仿-甲醇溶液从细胞中提取脂质,用蒸馏水分离非脂质提取物。提取的脂质在10ml圆烧瓶中干燥成薄膜,并通过涡流混合器(0.5%w/v溶液)用水剧烈摇动。然后超声10分钟,所得脂质在3000×g离心下4℃,取上清液,加5ml的PBS缓冲溶液溶解,于15,000-30,000×g,10-30min,1-6℃条件下高速离心,弃上清液。
4)取以上在第一次高转速的条件下离心得到的沉淀物;然后用PBS缓冲溶液溶解沉淀物,并以100,000-200,000×g,30-90min,1-6℃条件下超速离心,弃上清,收集沉淀B,即为所需提取的生物胞膜***。
实施例2
1)菌株培养:将嗜热栖热菌按1:100的比例接种到培养基中,利用梯度高热发酵法与高压、高渗透结合的方法,提高了多胺复合物、类胡萝卜素的产率。加NaCl至浓度为1.35%,调节发酵液渗透压至460mOsmol/kg,采用60-70℃,70-75℃,75-90℃每8H逐级升温,在终条件下,继续培养24h。
2)破壁:发酵后在4,000rpm,30min,4℃条件下离心收集菌体;用匀浆缓冲液(20mmol/L Tris-Cl pH8.0,100mmol/L NaCl,2mmol/L MgCl2,1mmol/L DTT)重悬菌体,在6,000rpm,10min,4℃条件下离心去上清;加入匀浆缓冲液重悬沉淀(约1g加入10ml缓冲液),再加入终浓度为1mmol/L的PMSF,冰浴超声破碎30min(振幅为55%,超声5s,停8s),得到细胞裂解液。
3)采用二次密度梯度离心法和超速离心法进行提取操作。
具体地,密度梯度离心法包括以下的步骤:将获得的细胞裂解液加入到质量百分比浓度分别为20%、30%、40%、50%、60%、70%的蔗糖溶液中重新悬浮得到重悬液,以150,000-300,000×g,60-90min,1-6℃条件下超速离心,收集蔗糖溶液的质量百分比浓度为40%-50%区域的液体C。将液体C加PBS缓冲溶液稀释,以100,000-200,000×g,30-90min,1-6℃条件下超速离心,弃上清,收集沉淀,即为所需提取的生物胞膜***。
表2不同蔗糖浓度时密度梯度离心(二次)的所得生物胞膜***囊泡对比
由表2可知,质量百分比浓度为40%-50%的蔗糖溶液中,超氧化物歧化酶(SOD)等生物酶活性最强。
实施例3
与实施例1的不同之处在于,步骤1)菌株培养:将嗜热栖热菌按1:100的比例接种到培养基中,在DO溶氧值小于15%(控制通氧)条件下,用500lx日光灯照射,从65℃开始,每小时上升2℃梯度发酵,在终条件下(75℃)继续培养至24小时时结束,收集菌体。
实施例4
与实施例3的不同之处在于,步骤1)菌株培养时,采用日光灯梯度照射法:0-2h用500lx,2-8h调整至1000lx,8h以后全程改为1500lx,直至24h发酵结束。
表3光照与未光照的生物胞膜***的类胡卜素及多胺含量对比
指标 | 未光照组 | 实施例3(500lx) | 实施例4(梯度照射法) |
总多胺(mg/g) | 6.19±0.68 | 39.18±1.76 | 52.43±3.12 |
类胡萝卜素(ppm) | 198.0±22.0 | 305.5±18.4 | 299.2±10.1 |
Mn2+(ppm) | 0.54±0.08 | 3.82±0.13 | 5.14±0.18 |
SOD(U/g) | 39.2±2.5 | 125.2±4.9 | 133.62±7.8 |
Fe3+(ppm) | 1.58±0.19 | 13.34±0.37 | 14.58±0.21 |
从表3可得,采用500lx和梯度照射法的光照组均可以有效增加总多胺、类胡萝卜素、SOD以及Mn2+和Fe3+金属离子的含量,其中,500lx条件下生物胞膜***中的总多胺含量提升了5倍,类胡萝卜素含量提升了52.29%。
实施例5
与实施例1的不同之处在于,步骤1)中采用3.4%的CaCl2高渗溶液,调节发酵液渗透压至600mOsmol/kg。
实施例6
与实施例1的不同之处在于,步骤1)发酵液采用等渗透压。
图1是不同生物胞膜***的总蛋白质电泳结果图,M-1是3.4%的CaCl2法所得的生物胞膜***(实施例5),M-2是等渗法所得的生物胞膜***(实施例6),M-3是500lx光照发酵所得生物胞膜***(实施例3),M-4是卵磷脂。
实施例7
与实施例1的不同之处在于,步骤1)中采用9%的CaCl2高渗溶液,调节发酵液渗透压至1500mOsmol/kg。
实施例8
与实施例1的不同之处在于,步骤1)中采用10%的甘油高渗溶液,调节发酵液渗透压至2100mOsmol/kg。
表4渗透压和渗透压调节溶质的影响
从表4可知,渗透压越高,菌株的耐高渗透压应变力越高,产生的多胺复合物和类胡萝卜素越多,而且糖脂的含量越少。
实施例9
与实施例2的不同之处在于:重悬介质为硅溶胶(Percoll)时:在4000rpm下低温离心,1小时,取上清液,按等体积加于密度梯度比重的Percoll密度梯度分层液上,室温下离心,1200r/min,25min;将每一层细胞分别吸出,各放入1只试管内,加PBS重悬液在4℃下洗涤3次,每次4000r/min离心10min,弃上清,即得所需提取的生物胞膜***。
所得的生物胞膜***中总多胺为31.43±0.86mg/g,类胡萝卜素为248.0±25.1ppm,糖脂为67.3±10.9ppm。对比表4中实施例2中的总多胺和类胡萝卜素的含量可知,采用Percoll重悬也可以得到与蔗糖溶液重悬时的效果。
实施例10
按实施例4(光照工艺)制得的生物胞膜***,对比(无光照,且等渗)但其他工艺条件相同下所得的生物胞膜***在光损伤修复模型中进行测试。通过模拟的UVB光源照射HaCaT细胞建立光损伤修复模型后加入受试物,检测受试物对光损伤细胞的修复能力。收集对数期细胞,用含10%FBS、1%SK的RPMI-1640培养液制成HaCaT细胞悬液,为5×104个/mL接种100μL/孔在96孔板中,于37℃、5%(v/v)CO2下培养,板四周孔用无菌PBS填充封边。细胞培养24h后,吸出培养液,用PBS洗一次,吸出培养液,再加入20uL的PBS缓冲溶液,进行UVB光源照射(UVB照射时间:UVB灯,开灯10分钟后用紫外线辐射强度1.05mW/cm2。辐射剂量控制为30mJ/cm2,辐射时间(30s),加入不同体积分数(0.5%、1%、3%、5%)的样品,以100ppm***为阳性对照组,以不含加药样品的培养液为对照组。继续培养24小时,进行MTT实验。MTT实验是一种检测细胞存活和生长的方法,其检测原理为活细胞线粒体中的琥珀酸脱氢酶能使外源性MTT还原为水不溶性的蓝紫色结晶甲瓒(Formazan)并沉积在细胞中,而死细胞无此功能。
记录以细胞存活率来评价样品对光损伤的修复能力,请参考附图2,实施例3制得的生物胞膜***具有更好的光损伤修复能力。
实施例11
以实施例1所制得的生物胞膜***,并选择不同体积分数(1%、3%、5%、10%、15%、20%、25%)为样品,50ppm的氢化可的松作为阳性对照组,LPS为对照组,在RAW细胞(NO)通过LPS诱导产生NO的消炎能力模型中评估。从附图3可知,实施例1制得的生物胞膜***随着浓度的升高,消炎能力不断增强,当生物胞膜***的体积分数达20%时,消炎能力增长逐渐放缓,当生物胞膜***的体积分数为25%时,可达到与50ppm氢化可的松接近的消炎能力。由此可得,本发明实施例1制得的生物胞膜***具有显著的消炎能力。
以上所述仅为本发明的优选实施例,并非因此即限制本发明的专利保护范围,凡是运用本发明说明书内容所作的等效结构变换,直接或间接运用在其他相关的技术领域,均同理包括在本发明的保护范围内。
Claims (7)
1.一种生物胞膜***的制备方法,其特征在于,具体包括以下步骤:
1)通过温度梯度法、调节光照强度、调节渗透压中的任一种或多种组合的方法在无糖培养基中进行菌株的高代谢培养;
2)收集菌体,采用匀浆缓冲液重悬菌体,离心,取沉淀的菌体进行破碎,得到细胞裂解液;
3)在细胞裂解液中加入氯仿-甲醇溶液提取脂质,将提取的脂质干燥成薄膜,再将薄膜加水混匀后进行离心,弃沉淀,收集上清液A;
4)对上清液A进行超速离心,弃上清液,收集沉淀B,得到所需的生物胞膜***;
其中,所述菌株为嗜热栖热菌,保藏编号为ATCC27634;
其中,所述温度梯度法采用60-90℃的温度范围内采用逐步升温的方式培养24h;所述调节光照强度采用单一光照强度或梯度光照强度,光照强度的调节范围为500-1500lx,光照时间为24-72h;所述调节渗透压通过添加渗透压调节溶质调节发酵液的渗透压至350-3100mOsmol/kg,所述渗透压调节溶质为盐或甘油。
2.一种生物胞膜***的制备方法,其特征在于,具体包括以下步骤:
1)通过温度梯度法、调节光照强度、调节渗透压中的任一种或多种组合的方法在无糖培养基中进行菌株的高代谢培养;
2)收集菌体,采用匀浆缓冲液重悬菌体,离心,取沉淀的菌体进行破碎,得到细胞裂解液;
3)将细胞裂解液加入重悬介质中得到重悬液,通过超速离心和密度梯度离心对分层的重悬液中生物胞膜***进行提纯,得到所需的生物胞膜***;
其中,所述菌株为嗜热栖热菌,保藏编号为ATCC27634;
其中,所述温度梯度法采用60-90℃的温度范围内采用逐步升温的方式培养24h;所述调节光照强度采用单一光照强度或梯度光照强度,光照强度的调节范围为500-1500lx,光照时间为24-72h;所述调节渗透压通过添加渗透压调节溶质调节发酵液的渗透压至350-3100mOsmol/kg,所述渗透压调节溶质为盐或甘油。
3.根据权利要求1或2所述的生物胞膜***的制备方法,其特征在于,所述生物胞膜***包括磷脂双分子层以及接枝于所述磷脂双分子层的多胺复合物和镶嵌于所述磷脂双分子层上的聚异戊二烯类化合物,所述多胺复合物包括任意一种或多种:
双端分别具有两个氨基的直链或支链烷基胺,
双端分别具有至少两个氨基且包含1-4个仲胺的直链或支链烷基胺,
一端具有至少一个氨基另一端为咪唑基的直链或支链烷基胺,
环状多胺;
所述聚异戊二烯类化合物选自四萜化合物、四萜化合物脂肪酸酯衍生物中的任意一种或多种的组合;
所述生物胞膜***中糖脂的含量≤100ppm。
4.根据权利要求3所述的生物胞膜***的制备方法,其特征在于,所述生物胞膜***还包括具有生物活性的氧化还原酶,所述氧化还原酶的活性中心具有金属离子;所述金属离子来自Mn、Ca、Mg、Zn、Cu、Fe中的任意一种或多种。
5.根据权利要求4所述的生物胞膜***的制备方法,其特征在于,所述氧化还原酶为超氧化物歧化酶,且其活性中心具有锰离子。
6.根据权利要求3所述的生物胞膜***的制备方法,其特征在于,所述多胺复合物为组胺、胍丁胺、戊二胺、精脒、腐胺、亚精胺、去甲精胺、高精胺、精胺、细胞色素c中的一种或多种的组合。
7.根据权利要求2所述的生物胞膜***的制备方法,其特征在于,所述重悬介质由蔗糖、甘油、聚蔗糖、氯化铯、氯化钠、三碘苯甲酰葡萄糖胺、硅溶胶、氟代碳中的至少一种配制而成。
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