CN108854599A - 一种基于交联溶菌酶的透析膜及其应用 - Google Patents
一种基于交联溶菌酶的透析膜及其应用 Download PDFInfo
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Abstract
本发明公开了一种基于交联溶菌酶的透析膜及其应用，该透析膜是溶菌酶相转变形成的二维纳米薄膜用戊二醛交联后粘附在PET核孔膜上，形成的以交联溶菌酶纳米薄膜为致密皮层、PET核孔膜为支撑层的透析膜。本发明透析膜易实现大面积制备，同时又具有低成本、低能耗、环保等特点，避免了传统聚合物膜合成过程中的步骤繁琐及环境污染问题，且透析膜具有较好的生物相容性，对不同分子具有大小选择性分离，能够完全截留3.2nm以上的分子，对小分子有较快的扩散速率，可作为血液净化膜，清除尿素、肌酸肝及中大分子毒素和硫酸吲哚酚，是一种较为理想的透析膜材料。
Description
技术领域
本发明涉及一种透析膜，具体涉及一种制备简单、绿色、温和且孔径可控的透析膜，以及该透析膜的应用。
背景技术
膜分离技术由于具有分离效率高、无二次污染及操作简单等特点，目前已广泛运用于石油化工、污水处理、医药卫生、食品加工等领域。膜材料是膜分离技术的核心内容之一，目前常用的膜材料主要为有机合成高分子材料，然而聚合物膜的制备方法复杂，通常容易吸附原料液中的有机物质(如蛋白质、胶体和微生物等)；在与血液接触时，膜表面的蛋白质非特异性吸附还会引起凝血等负面效应，造成分离膜的渗透量降低，分离特异性降低。血液透析治疗是目前尿毒症患者常用的治疗手段之一，血液净化膜主要通过扩散/对流原理进行物质交换，并达到清除体内的代谢废物，维持电解质和清除体内过多的水分为目的。而常规血液透析膜无法达到对中大分子毒素的清除。因此，开发具有良好分离性能的同时，还具有简单易行，价格低廉的分离膜具有重要的现实意义。
利用纳米结构材料自组装制备的选择性分离膜是近年来分离膜领域的一个新的尝试。时至今日，对于工业应用来说，具有选择透过性的大部分纳米复合膜并不能大面积生产。
发明内容
本发明所要解决的技术问题在于针对上述现有技术的缺陷，提供一种基于交联溶菌酶的透析膜，以及该透析膜的应用。
解决上述技术问题所采用的基于交联溶菌酶的透析膜是：将溶菌酶相转变形成的溶菌酶二维纳米薄膜用戊二醛交联后粘附在PET核孔膜上，形成以交联溶菌酶纳米薄膜为致密皮层、PET核孔膜为支撑层的透析膜。
上述溶菌酶相转变形成的溶菌酶二维纳米薄膜的方法为：将10～100mmol/L三(2-羧乙基)膦的4-羟乙基哌嗪乙磺酸缓冲溶液用NaOH调节至pH值为6.0～8.0，然后将其与1～30mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液等体积混合后直接铺满基材表面，室温培育2～6小时，在基材上液体的气液界面形成一层溶菌酶二维纳米薄膜。
上述溶菌酶相转变形成的溶菌酶二维纳米薄膜用戊二醛交联的方法为：将溶菌酶二维纳米薄膜转移至质量分数为0.2％～2％的戊二醛水溶液中，室温交联2～6小时。
上述PET核孔膜的膜直径为25mm、厚度为12μm、孔径大小为1～10μm。
本发明基于交联溶菌酶的透析膜在混合蛋白质分离中的应用，其中所述的混合蛋白质为牛血清蛋白和胰岛素，或者肌红蛋白和胰岛素。
本发明基于交联溶菌酶的透析膜在混合染料分离中的应用，其中所述的混合染料为甲基蓝和甲基橙，或者甲基蓝和罗丹明B。
本发明基于交联溶菌酶的透析膜在去除尿毒素中的应用，其中所述的尿毒素为尿素、肌酸肝、β-微球蛋白以及硫酸吲哚酚。
本发明的有益效果如下：
1、本发明透析膜由溶菌酶的寡聚物密堆积而成，无色透明，为纯蛋白质膜，根据溶菌酶的浓度可以控制膜的厚度，且随膜厚度的增加其孔大小从3.4nm减小至1.8nm。
2、本发明透析膜具有较好的生物相容性，可作为血液净化膜，能够清除尿素、肌酸肝及中大分子毒素和硫酸吲哚酚，且对于大分子蛋白具有更高的截留率，是一种较为理想的透析膜材料，其对分子大小在3.2nm以上的蛋白质的截留率≥98％、对尿素及肌酸肝的清除率分别为82.2％和81.3％，对较难清除的蛋白质结合毒素的清除率为33.1％。
3、本发明透析膜能够有效分离牛血清蛋白和胰岛素、肌红蛋白和胰岛素、甲基蓝和甲基橙、甲基蓝和罗丹明B。
4、本发明透析膜制备方法简单，易实现大面积制备，同时又具有低成本、低能耗、环保等特点，避免了传统聚合物膜合成过程中的步骤繁琐及环境污染问题。
附图说明
图1是实施例1中交联溶菌酶纳米薄膜的扫描电镜图。
图2是实施例1中交联溶菌酶纳米薄膜的透射电镜平面图。
图3是实施例1中交联溶菌酶纳米薄膜的原子力显微镜图。
图4是不同厚度的交联溶菌酶纳米薄膜的孔径分布。
图5是实施例1得到的透析膜对尿素、硫酸吲哚酚、肌酸肝及β-微球蛋白的清除。
具体实施方式
下面结合附图和实施例对本发明进一步详细说明，但本发明的保护范围不仅限于这些实施例。
实施例1
将60μL 50mmol/L三(2-羧乙基)膦的4-羟乙基哌嗪乙磺酸缓冲溶液用NaOH调节至pH值为7.0，然后将其与60μL 2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液混合均匀后直接铺满在18mm×18mm的盖玻片表面，室温培育2小时，在盖玻片上液体的气液界面形成一层溶菌酶二维纳米薄膜；将气液界面的溶菌酶二维纳米薄膜转移至质量分数为1％的戊二醛水溶液中，室温交联2小时后，得到交联溶菌酶纳米薄膜。
将上述交联溶菌酶纳米薄膜粘附在膜直径为25mm、厚度为12μm、孔径大小为10μm的PET核孔膜上，得到以交联溶菌酶纳米薄膜为致密皮层、PET核孔膜为支撑层的透析膜。
实施例2
本实施例中，用等体积4mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液替换实施例1中2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液，其他步骤与实施例1相同，得到透析膜。
实施例3
本实施例中，用等体积6mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液替换实施例1中2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液，其他步骤与实施例1相同，得到透析膜。
实施例4
本实施例中，用等体积8mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液替换实施例1中2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液，其他步骤与实施例1相同，得到透析膜。
实施例5
本实施例中，用等体积10mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液替换实施例1中2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液，其他步骤与实施例1相同，得到透析膜。
实施例6
本实施例中，用等体积20mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液替换实施例1中2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液，其他步骤与实施例1相同，得到透析膜。
实施例7
本实施例中，用等体积30mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液替换实施例1中2mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液，其他步骤与实施例1相同，得到透析膜。
发明人对实施例1～7中制备的交联溶菌酶纳米薄膜进行扫描电镜表征，结果显示其对应的膜厚度依次为50(见图1)、60、90、100、120、220和250nm，该纳米薄膜是由溶菌酶相转变后的20nm大小的寡聚物密堆积而成(如图2、3)。将制备的交联溶菌酶纳米薄膜用孔径为6mm的PET对其面积进行控制，将该膜漂浮于5mL超纯水中，且在该膜上滴加50μL 0.5mg/mL不同分子量的PEG，采用紫外可见吸收光谱对静置24小时前后的溶液进行透过性监控，并分析膜的孔径大小分布，结果如图4所示。通过拟合膜对不同分子量的截留率得到膜的孔径大小分布，随着膜厚度的增加膜的孔径从3.4nm减小至1.8nm。
实施例8
实施例1的透析膜在分离甲基蓝和甲基橙、甲基蓝和罗丹明B、肌红蛋白和胰岛素、牛血清蛋白和胰岛素中的应用，具体方法如下：
将透析膜漂浮于5mL超纯水中，分别在该膜上滴加50μL含50mg/L甲基蓝和50mg/L甲基橙的水溶液、50μL含50mg/L甲基蓝和50mg/L罗丹明的水溶液、50μL含10g/L肌红蛋白和10g/L胰岛素的水溶液、50μL含10g/L肌红蛋白和10g/L胰岛素的水溶液、50μL含10g/L牛血清蛋白和10g/L胰岛素的水溶液，室温静置24小时后，采用紫外可见吸收光谱对溶液进行透过性监控。实验结果显示，该透析膜能够完全通过分子直径小于2nm的分子，分子直径大于3nm并且带有负电荷的甲基蓝、牛血清蛋白和肌红蛋白能够被完全截留，而甲基橙的截留率仅为0.3％、罗丹明B的截留率为1.4％、胰岛素的截留率为22.6％，其中甲基橙的扩散速率达到606nmolcm-2h-1，罗丹明B的扩散速率为308nmol cm-2h-1，能够实现甲基蓝和甲基橙、甲基蓝和罗丹明、肌红蛋白和胰岛素、牛血清蛋白和胰岛素的快速、有效分离。
实施例9
实施例1的透析膜在去除尿素、肌酸肝、β-微球蛋白以及硫酸吲哚酚中的应用，具体方法如下：
模拟液是含有1mg/mL牛血清蛋白和25mg/L硫酸吲哚酚、40mg/L β-微球蛋白、100mg/L肌酸肝和1.5mg/mL尿素的水溶液，模拟液及透析液(水)的流速都为10mL/min，用透析膜透析4小时。测试透析膜对牛血清蛋白和硫酸吲哚酚、β-微球蛋白、肌酸肝、尿素的清除率，如图5所示，该透析膜能够截留大分子牛血清蛋白，对硫酸吲哚酚、尿素、β-微球蛋白及肌酸肝的清除效果较好，清除率依次为33.1％、82.2％、50.3％和81.3％。
Claims (7)
1.一种基于交联溶菌酶的透析膜，其特征在于：该透析膜是溶菌酶相转变形成的二维纳米薄膜用戊二醛交联后粘附在PET核孔膜上，形成的以交联溶菌酶纳米薄膜为致密皮层、PET核孔膜为支撑层的透析膜。
2.根据权利要求1所述的基于交联溶菌酶的透析膜，其特征在于所述溶菌酶相转变形成溶菌酶二维纳米薄膜的方法为：将10～100mmol/L三(2-羧乙基)膦的4-羟乙基哌嗪乙磺酸缓冲溶液用NaOH调节至pH值为6.0～8.0，然后将其与1～30mg/mL溶菌酶的4-羟乙基哌嗪乙磺酸缓冲溶液等体积混合后直接铺满基材表面，室温培育2～6小时，在基材上液体的气液界面形成一层溶菌酶二维纳米薄膜。
3.根据权利要求1所述的基于交联溶菌酶的透析膜，其特征在于所述溶菌酶相转变形成的溶菌酶二维纳米薄膜用戊二醛交联的方法为：将溶菌酶二维纳米薄膜转移至质量分数为0.2％～2％的戊二醛水溶液中，室温交联2～6小时。
4.根据权利要求1所述的基于交联溶菌酶的透析膜，其特征在于：所述PET核孔膜的膜直径为25mm、厚度为12μm、孔径大小为1～10μm。
5.权利要求1所述的基于交联溶菌酶的透析膜在混合蛋白质分离中的应用，所述的混合蛋白质为牛血清蛋白和胰岛素，或者肌红蛋白和胰岛素。
6.权利要求1所述的基于交联溶菌酶的透析膜在混合染料分离中的应用，所述的混合染料为甲基蓝和甲基橙，或者甲基蓝和罗丹明B。
7.权利要求1所述的基于交联溶菌酶的透析膜在去除尿毒素中的应用，所述的尿毒素为尿素、肌酸肝、β-微球蛋白以及硫酸吲哚酚。
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| CN108854599B (zh) * | 2018-05-14 | 2020-10-16 | 陕西师范大学 | 一种基于交联溶菌酶的透析膜及其应用 |
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- 2019-05-07 WO PCT/CN2019/085848 patent/WO2019218898A1/zh active Application Filing
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| WO2019218898A1 (zh) * | 2018-05-14 | 2019-11-21 | 陕西师范大学 | 一种基于交联蛋白质的分离膜及其应用 |
| US12011694B2 (en) | 2018-05-14 | 2024-06-18 | Shaanxi Normal University | Crosslinked protein-based separation membrane and application thereof |
| CN109748360A (zh) * | 2019-02-27 | 2019-05-14 | 南京大学 | 一种多功能杂化超滤膜、制备方法及应用 |
| CN110975826A (zh) * | 2019-05-30 | 2020-04-10 | 陕西师范大学 | 一种基于交联蛋白质的吸附材料及其回收贵金属的应用 |
| WO2020238505A1 (zh) * | 2019-05-30 | 2020-12-03 | 陕西师范大学 | 一种基于交联蛋白质的吸附材料及其回收贵金属的应用 |
| CN110975826B (zh) * | 2019-05-30 | 2022-03-22 | 陕西师范大学 | 一种基于交联蛋白质的吸附材料及其回收贵金属的应用 |
| CN110563979B (zh) * | 2019-09-23 | 2022-10-21 | 陕西师范大学 | 一种基于巯基与二硫键交换反应的蛋白质纳米薄膜及其应用 |
| CN110563979A (zh) * | 2019-09-23 | 2019-12-13 | 陕西师范大学 | 一种基于巯基与二硫键交换反应的蛋白质纳米薄膜及其应用 |
| CN111603942A (zh) * | 2020-06-12 | 2020-09-01 | 吉林大学 | 一种基于界面组装方式的蛋白分离膜、制备方法及其应用 |
| CN112574578A (zh) * | 2020-11-19 | 2021-03-30 | 陕西师范大学 | 蛋白质/多糖复合纳米薄膜及其防止导电涂层产生裂纹的应用 |
| CN112892226A (zh) * | 2021-01-29 | 2021-06-04 | 武汉大学 | 一种蛋白纳米纤维素复合多功能分离膜、制备方法及其应用 |
| CN115216034A (zh) * | 2021-04-16 | 2022-10-21 | 复旦大学 | 丝蛋白/溶菌酶可转移蛋白质纳米薄膜或涂层及其制备 |
| CN115216034B (zh) * | 2021-04-16 | 2023-08-29 | 复旦大学 | 丝蛋白/溶菌酶可转移蛋白质纳米薄膜或涂层及其制备 |
| CN115400604A (zh) * | 2022-09-06 | 2022-11-29 | 西北工业大学 | 一种用于镁锂分离的正电荷纳滤膜涂层及制备方法 |
| CN115400604B (zh) * | 2022-09-06 | 2023-08-18 | 西北工业大学 | 一种用于镁锂分离的正电荷纳滤膜涂层及制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| US20210187448A1 (en) | 2021-06-24 |
| US12011694B2 (en) | 2024-06-18 |
| WO2019218898A1 (zh) | 2019-11-21 |
| CN108854599B (zh) | 2020-10-16 |
| US20220134294A1 (en) | 2022-05-05 |
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