CN106335234B - 一种基于非共价修饰的石墨烯蛋白复合薄膜及制备方法 - Google Patents

一种基于非共价修饰的石墨烯蛋白复合薄膜及制备方法 Download PDF

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CN106335234B
CN106335234B CN201610694149.3A CN201610694149A CN106335234B CN 106335234 B CN106335234 B CN 106335234B CN 201610694149 A CN201610694149 A CN 201610694149A CN 106335234 B CN106335234 B CN 106335234B
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毛红菊
周麟
马云飞
金庆辉
赵建龙
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Shanghai Institute of Microsystem and Information Technology of CAS
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Abstract

本发明涉及一种基于非共价修饰的石墨烯蛋白复合薄膜及制备方法，所述复合薄膜包括二氧化硅衬底、单层石墨烯和蛋白薄膜。制备方法包括：将通过化学气相沉积生长的单层石墨烯薄膜转移至二氧化硅衬底的表面，将石墨烯薄膜表面的光刻胶除去后在二氧化硅衬底表面留下单层石墨烯；将蛋白溶液滴加到单层石墨烯表面在60～90℃热失活处理1～10min，在单层石墨烯表面形成蛋白薄膜，即得。本发明通过热失活方法在石墨烯表面实现蛋白薄膜的非共价的修饰，石墨烯‑蛋白薄膜厚度可以控制在纳米级，薄膜均匀，操作简单，易于实现；蛋白薄膜表面具有氨基和羧基，可以作为后续生物分子诊断的平台，具有良好的应用前景。

Description

一种基于非共价修饰的石墨烯蛋白复合薄膜及制备方法
技术领域
本发明属于石墨烯生物传感器领域，特别涉及一种基于非共价修饰的石墨烯蛋白复合薄膜及制备方法。
背景技术
石墨烯是一种具有单层碳原子结构的二维材料，鉴于其稳定的化学结构以及优异的光学、机械和电学特性，使得石墨烯成为纳米生物电学检测领域中一种理想的材料。石墨烯器件被用于蛋白、DNA、神经细胞动作电位以及细菌的检测。而在石墨烯生物传感器的构建过程中，石墨烯表面的生物分子修饰是其中至关重要的环节。石墨烯表面的修饰方法通常可以分为共价修饰和非共价修饰。石墨烯的共价修饰通常比较稳定但是会损坏石墨烯的电学性能，而非共价修饰通常可以有效地偶联生物分子而最大限度地保留石墨烯原有的电学特性。因此石墨烯的非共价修饰是生物传感器的构建中的理想选择。
石墨烯的非共价修饰主要是通过π堆叠作用和疏水作用来实现的，π堆叠作用主要通芳香环与石墨烯之间的吸附来实现。通常的采用非共价修饰的双功能化学分子，一端吸附在石墨烯的表面，另一端进行生物探针的偶联。其中典型的偶联分子，如1-pyrenebutanoic acid succinimidyl ester、NHS ester tripod、肽链以及小牛血清(bovine serum albumin，BSA)等。1-pyrenebutanoic acid succinimidyl ester和NHSester tripod偶联分子对光与湿度较为敏感，肽链与BSA在石墨烯表面的吸附量较少，而且通常偶联分子的吸附量无法控制，很难满足石墨烯应用于生物传感器中的实际需求。因此石墨烯表面均匀、可控和稳定的修饰对石墨烯生物传感器的应用具有重要的意义。
发明内容
本发明所要解决的技术问题是提供一种基于非共价修饰的石墨烯蛋白复合薄膜及制备方法，通过热失活方法在石墨烯表面实现蛋白薄膜的非共价的修饰，石墨烯-蛋白薄膜厚度可以控制在纳米级，薄膜均匀，操作简单，易于实现；蛋白薄膜表面具有氨基和羧基，可以作为后续生物分子诊断的平台，具有良好的应用前景。
本发明的一种基于非共价修饰的石墨烯蛋白复合薄膜，所述复合薄膜包括二氧化硅衬底、单层石墨烯和蛋白薄膜；所述单层石墨烯位于二氧化硅衬底表面，单层石墨烯表面具有非共价修饰的蛋白薄膜，所述蛋白薄膜表面带有氨基与羧基。
所述蛋白薄膜为牛血清蛋白(bovine serum albumin，BSA)。
所述蛋白薄膜通过热失活吸附在石墨烯表面。
所述蛋白薄膜厚度为纳米级别。
本发明的一种基于非共价修饰的石墨烯蛋白复合薄膜的制备方法，包括：
将通过化学气相沉积生长的单层石墨烯薄膜转移至二氧化硅衬底的表面，将石墨烯薄膜表面的光刻胶除去后在二氧化硅衬底表面留下单层石墨烯；将浓度为0.18μM～1.8mM的蛋白溶液滴加到单层石墨烯表面在60～90℃热失活处理1～10min，在单层石墨烯表面形成蛋白薄膜，即得非共价修饰的石墨烯蛋白复合薄膜。
所述蛋白溶液的溶剂为磷酸盐缓冲液(PBS溶液)、邻苯二甲酸氢钾缓冲溶液或硼酸盐缓冲液。
制备石墨烯蛋白复合薄膜的方法还包括：
(1)配制不同的BSA溶液：用去离子水配制不同浓度的BSA溶度，采用不同离子浓度的溶剂配制相同浓度的BSA溶液，用相同浓度的PBS(pH7.0)、邻苯二甲酸氢钾缓冲溶液(pH3.5)以及硼酸盐缓冲液(pH9.0)分别作为不同pH值的缓冲液；
(2)用相同浓度溶解在去离子水中的BSA溶液滴加在石墨烯表面进行失活，控制不同的失活时间在石墨烯表面形成不同厚度的薄膜；
(3)将溶解在去离子水中不同浓度的BSA溶液滴加在石墨烯表面进行失活，在相同的温度下失活相同的时间，可以在石墨烯表面形成不同厚度的薄膜；
(4)用不同离子浓度的溶剂配制相同浓度的BSA溶液，在相同的温度和时间下失活，可以在石墨烯表面形成不同厚度的薄膜；
(5)用相同浓度的PBS、邻苯二甲酸氢钾缓冲溶液以及硼酸盐缓冲液配制相同浓度的BSA溶液，滴加在石墨烯表面进行失活，实现对石墨烯的不同掺杂。
本发明通过热失活方法，通过控制相同溶剂中BSA溶液的浓度、以及相同浓度的BSA溶液溶剂中的离子浓度，以及失活的时间，可以在石墨烯表面形成不同纳米级厚度的石墨烯-蛋白薄膜，通过控制溶解BSA溶液的pH值可以实现对石墨烯不同电荷类型的掺杂，通过表征可以确定该薄膜表面具有氨基和羧基，可以作为后续生物分子诊断的平台。
有益效果
本发明通过热失活方法在石墨烯表面实现蛋白薄膜的非共价的修饰，石墨烯-蛋白薄膜厚度可以控制在纳米级，薄膜均匀，操作简单，易于实现；蛋白薄膜表面具有氨基和羧基，可以作为后续生物分子诊断的平台，具有良好的应用前景。
附图说明
图1为本发明石墨烯蛋白复合薄膜的结构示意图；其中，1为二氧化硅衬底，2为单层石墨烯，3为蛋白薄膜；
图2a-d为本发明石墨烯蛋白复合薄膜表面基团的X射线光电子能谱图。
具体实施方式
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。此外应理解，在阅读了本发明讲授的内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
实施例1
如图1所示为二氧化硅衬底上单层石墨烯表面形成不同纳米级厚度的蛋白薄膜结构示意图。将通过化学气相沉积生长的单层石墨烯薄膜转移至二氧化硅衬底的表面，将石墨烯薄膜表面的光刻胶除去后在二氧化硅衬底表面留下单层石墨烯；将蛋白溶液滴加到单层石墨烯热失活处理，在单层石墨烯表面形成蛋白薄膜，即得非共价修饰的石墨烯蛋白复合薄膜。
将浓度1.8mM溶解在去离子水中的BSA溶液，将加热失活的温度设定在75℃，控制蛋白失活的时间分别为2min、4min和6min，可在石墨烯表面形成不同纳米级厚度的蛋白薄膜，石墨烯表面蛋白薄膜的厚度与失活时间呈正相关。
将溶解在去离子水中浓度为1.8mM、18uM和0.18uM的BSA溶液滴加在石墨烯表面进行失活，在失活温度为75℃下失活4min，可以在石墨烯表面形成不同厚度的薄膜，石墨烯表面蛋白薄膜的厚度与BSA溶液的浓度呈正相关。
将1.8mM的BSA溶液分别溶解在10mM的PBS溶液、1mM的PBS溶液以及去离子水中，然后滴加在石墨烯表面进行失活，在失活温度为75℃下失活4min，可以在石墨烯表面形成不同厚度的薄膜，石墨烯表面蛋白薄膜的厚度与溶解BSA的溶剂中的离子浓度呈负相关。
将1.8mM的BSA溶液分别溶解在1mM PBS溶液(pH7.0)、1mM邻苯二甲酸氢钾缓冲溶液(pH3.5)以及1mM硼酸盐缓冲液(pH9.0)中，然后滴加在石墨烯表面进行失活，在失活温度为75℃下失活4min，蛋白BSA的等电点在5左右，在pH值偏离蛋白等电点的溶液中，由于蛋白分子电负性的不同可对石墨烯形成不同类型的掺杂，使得石墨烯的狄拉克点往不同的方向偏移。利用X射线光电子能谱对石墨烯-蛋白薄膜的表面基团进行表征，如图2所示的石墨烯表面蛋白薄膜表面碳元素和氮元素的X射线光电子能谱图，蛋白薄膜表面的氨基和羧基可以从碳元素和氮元素的特征峰以及肩峰的位置得到证实。

Claims (5)

1.一种基于非共价修饰的石墨烯蛋白复合薄膜，其特征在于：所述复合薄膜包括二氧化硅衬底、单层石墨烯和蛋白薄膜；所述单层石墨烯位于二氧化硅衬底表面，单层石墨烯表面具有非共价修饰的蛋白薄膜，所述蛋白薄膜表面带有氨基与羧基；所述蛋白薄膜通过热失活吸附在石墨烯表面。
2.根据权利要求1所述的一种基于非共价修饰的石墨烯蛋白复合薄膜，其特征在于：所述蛋白薄膜为牛血清蛋白BSA。
3.根据权利要求1所述的一种基于非共价修饰的石墨烯蛋白复合薄膜，其特征在于：所述蛋白薄膜厚度为纳米级别。
4.一种基于非共价修饰的石墨烯蛋白复合薄膜的制备方法，包括：
将通过化学气相沉积生长的单层石墨烯薄膜转移至二氧化硅衬底的表面，将石墨烯薄膜表面的光刻胶除去后在二氧化硅衬底表面留下单层石墨烯；将浓度为0.18μM～1.8mM的蛋白溶液滴加到单层石墨烯表面在60～90℃热失活处理1～10min，在单层石墨烯表面形成蛋白薄膜，即得非共价修饰的石墨烯蛋白复合薄膜。
5.根据权利要求4所述的一种基于非共价修饰的石墨烯蛋白复合薄膜的制备方法，其特征在于：所述蛋白溶液的溶剂为磷酸盐缓冲液、邻苯二甲酸氢钾缓冲溶液或硼酸盐缓冲液。
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CN107167608B (zh) * 2017-04-06 2019-03-19 中国科学院上海微系统与信息技术研究所 一种基于多功能纳米级蛋白薄膜的石墨烯肿瘤标志物传感器及其制备方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102433304B (zh) * 2011-12-06 2014-04-16 中国科学院苏州纳米技术与纳米仿生研究所 用于细胞培养的石墨烯衬底及其制备方法
WO2014056896A2 (en) * 2012-10-08 2014-04-17 Albert-Ludwigs-Universität Freiburg One-step biomolecular immobilisation procedure and products thereof
CN103901089B (zh) * 2014-04-16 2016-08-24 国家纳米科学中心 检测神经细胞电生理信号的传感器及制作方法和检测方法
CN105460882B (zh) * 2015-12-04 2017-09-15 中国科学院上海微系统与信息技术研究所 一种石墨烯三维微电极阵列芯片、方法及其应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Harnessing Denatured Protein for Controllable Bipolar Doping of a Monolayer Graphene;Sung Kyu Jang;《ACS Applied Materials & Interfaces》;20150731;1250-1256 *
Investigation of Controllable Nanoscale Heat-Denatured Bovine;Lin Zhou;《Langmuir》;20161115;12626-12631 *
石墨烯的表面修饰、毒理学评价与体外降解行文研究;李英杰;《中国优秀硕士学位论文全文数据库 工程科技I辑》;20140930;B020-82 *

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