CN107817241B - 一种生物传感器、制备方法及检测乙酰胆碱酯酶的应用、一种检测有机磷农药的方法 - Google Patents
一种生物传感器、制备方法及检测乙酰胆碱酯酶的应用、一种检测有机磷农药的方法 Download PDFInfo
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Abstract
本发明提供了一种生物传感器、制备方法及检测乙酰胆碱酯酶的应用、一种检测有机磷农药的方法。利用乙酰胆碱具有模拟酶催化氧化TMB,使其变色,而乙酰胆碱酯酶能使乙酰胆碱分解失活,无法催化过氧化氢的分解,使得TMB溶液无法产生显色反应,进而构建吸光度与乙酰胆碱酯酶线性关系,实现对乙酰胆碱酯酶的定量检测。进一步的,有机磷农药能够有效的抑制乙酰胆碱酯酶的活性,从而导致乙酰胆碱酯酶失活,失活的乙酰胆碱酯酶无法使乙酰胆碱分解,进而使TMB溶液发生显色反应,实现对有机磷农药的检测。
Description
技术领域
本发明属于生物传感器领域,具体涉及一种生物传感器、制备方法及检测乙酰胆碱酯酶的应用、一种检测有机磷农药的方法。
背景技术
乙酰胆碱酯酶(AChE)作为一种生物催化剂的酶,在几乎所有的代谢过程中都起到重要的作用,如控制相关反应,提供用于细胞增殖和代谢所需的物质和能量。乙酰胆碱酯酶(AChE)作为初级胆碱酯酶,在神经系统中负责生物信号传输。它可以通过将乙酰胆碱水解成胆碱来特异性调节乙酰胆碱水平,从而导致在胆碱能突触的神经传递的终止。重要的是,许多神经退行性疾病,包括帕金森病,阿尔茨海默病和亨廷顿氏病与受损的AChE据研究有密切的关系,有证明抑制AChE活性可导致乙酰胆碱在突触中积累传递是疾病治疗的有效策略。此外,有机磷杀虫剂(OPs)对AChE活性的抑制也对人类健康十分有害。
因此,AChE活性的测量和监测具有巨大的现实意义和经济价值。在过去十多年来,已经探索了各种方法来监测AChE活性,包括比色Ellman测定和化学发光和电化学测定。然而,常规策略的某些缺点仍然存在,例如低检测灵敏度,假阳性效应,消耗时间和劳力,并且需要复杂的仪器,显然限制了这些应用方法。
显然,提高灵敏度并提高AChE检测的选择性是未来现场监测的需求方向。
发明内容
本发明的目的在于提供一种生物传感器及其制备方法。
本发明还提供了一种生物传感器检测乙酰胆碱酯酶的应用,利用乙酰胆碱具有模拟酶催化氧化TMB,使其变色,而乙酰胆碱酯酶能使乙酰胆碱分解失活,无法催化过氧化氢的分解,使得TMB溶液无法产生显色反应,进而构建吸光度与乙酰胆碱酯酶线性关系,实现对乙酰胆碱酯酶的定量检测。
本发明还提供了一种检测有机磷农药的方法,有机磷农药能够有效的抑制乙酰胆碱酯酶的活性,从而导致乙酰胆碱酯酶失活,失活的乙酰胆碱酯酶无法使乙酰胆碱分解,进而使TMB溶液发生显色反应,实现对有机磷农药的检测。
本发明提供的一种生物传感器的制备方法,包括以下步骤:
将乙酰胆碱缓冲溶液和乙酰胆碱酯酶缓冲溶液混合培养,将混合溶液加入含过氧化氢的TMB溶液中,即可。
所述混合培养是指35℃-40℃培养10-15min。
所述乙酰胆碱缓冲溶液和乙酰胆碱酯酶缓冲溶液配置所用的缓冲溶液是0.01M,pH=7.4的PBS缓冲溶液。
体系中TMB终浓度为2.5mg/ml,H2 O2终浓度为0.03%(V/V)。
本发明提供的一种生物传感器,采用上述方法制备得到。
本发明提供的一种生物传感器检测乙酰胆碱酯酶的应用,具体检测方法为:
将乙酰胆碱缓冲溶液和不同浓度的乙酰胆碱酯酶缓冲溶液混合培养,将混合溶液加入含过氧化氢的TMB溶液中,检测吸收波,构建吸光度与乙酰胆碱酯酶浓度的线性关系,进而实现对乙酰胆碱酯酶的检测。
所述混合培养是指35℃-40℃培养10-15min。
进一步的,乙酰胆碱终浓度为10mM/L,乙酰胆碱酯酶终浓度分别为0.2mU/mL、0.4mU/mL、0.6mU/mL、0.8mU/mL、1.0mU/mL、1.2mU/mL和1.4mU/mL。
进一步的,得到的线性的标准曲线方程A=-0.10885C+0.1937,相关系数R2=0.99729。
本发明提供的一种利用上述生物传感器检测检测有机磷农药的方法,具体检测方法为:
将乙酰胆碱酯酶缓冲溶液和分别与不同浓度的有机磷农药混合,得混合溶液,然后分别加入乙酰胆碱缓冲溶液中,最后将所得混合液分别加入到含有过氧化氢的TMB溶液中发生显色反应,检测吸收波,构建有机磷农药浓度与吸光度的线性关系,进而实现对有机磷农药的检测。
有机磷农药配制:取相应量的有机磷放入容量瓶,用三次水配制相应浓度的有机磷农药。
所述混合是指35℃--40℃反应5--10min。
所述乙酰胆碱酯酶终浓度为1.4mU/mL;所述的有机磷农药的终浓度为10mU/uL、20mU/uL、30mU/uL、40mU/uL、50mU/uL、60mU/uL、70mU/uL、80mU/uL、90mU/uL、100mU/uL、110mU/uL、120mU/uL、130mU/uL、140mU/uL;乙酰胆碱终浓度为10mM/L。
进一步的,所得线性关系标准曲线方程A=0.00189C+0.1679,相关系数R2=0.99323。
本发明提供了一种简便、快速、灵敏的比色法检测乙酰胆碱酯酶和有机磷农药的方法,利用乙酰胆碱具有模拟酶催化氧化TMB,使其变色。用于敏感检测AChE及其抑制剂,该项目具有较大的潜在可能。
附图说明
图1为基于比色法检测乙酰胆碱酯酶(AChE)及其抑制剂的方法及应用示意图;
图2A为乙酰胆碱具有模拟酶活性的可行性图(吸收波长652,时间720s);
A为TMB的空白对照;
B为加入10mM的胆碱;
C为加入10mM的乙酰胆碱;
图2B为乙酰胆碱模拟酶活性随其浓度增加在紫外可见下吸收波;
图3A为TMB浓度和其反应速率的关系图;
图3B为根据米氏方程计算其TMB和其速率计算方程图;
图3C为H2O2浓度和其反应速率的关系图;
图3D为根据米氏方程计算其TMB和其速率计算方程图;
图4A为pH值对本实验的影响图;
图4B为时间对本实验的优化图;
图4C为实验温度的优化图;
图5A为实验可行性图1,底物为含H2O2的TMB溶液;
a为加入乙酰胆碱的溶液;
b为加入乙酰胆碱和乙酰胆碱酯酶混合液的;
c为加入胆碱的溶液;
d为加入乙酰胆碱酯酶的溶液;
图5B为加入有机磷农药的可行性图,底物为含H2O2的TMB溶液;
a为加入乙酰胆碱和乙酰胆碱酯酶混合液的;
b为加入乙酰胆碱酯酶和有机磷农药反应后加入乙酰胆碱的溶液;
图6A为不同浓度乙酰胆碱酯酶的吸收波图;
图6B为在吸收波长652nm处不同浓度乙酰胆碱酯酶的标准曲线;
图7A为不同浓度有机磷农药的吸收波图;
图7B为在吸收波长652nm处不同浓度有机磷农药的标准曲线。
具体实施方式
乙酰胆碱具有模拟酶活性,能催化过氧化氢分解,从而导致TMB被氧化显色。进行一系列实验探究,发现如图2A所示,有乙酰胆碱能使TMB从无色变成蓝色,而空白组和胆碱组都无法使TMB变色。进一步研究中,我们发现改变乙酰胆碱的浓度,其导致的TMB颜色的深度也是不一样的。所以得出TMB显色最大吸光度的大小是随乙酰胆碱浓度的变化而变化。取配制好三组的含过氧化氢的TMB溶液在试管中,第一组空白,另外一组加入0.01mM/L乙酰胆碱,另一组0.01mM/L胆碱,发现只有加入乙酰胆碱的TMB溶液由原来的无色变成蓝色。证明本实验的可行性。
进一步探究乙酰胆碱使含过氧化氢溶液的3,3',5,5'-四甲基联苯胺(TMB)变色,确定一系列优化条件:
取配制好的含过氧化氢的TMB溶液在试管中,加入0.01mM/L乙酰胆碱,改变反应的条件,分三组实验。第一组乙酰胆碱溶液把pH分别调整为4,5,6,7,7.4,8,9,其他实验条件不变,加入相同的含过氧化氢的TMB溶液中,发现pH为7的时吸光度最大。
第二组,只改变温度从20,25,30,35,37,40,45,50℃,其他实验条件不变,温度为37℃时其吸光度最大。
第三组,只改变反应时间0-720秒,时间在720秒后最大吸光度趋于稳定。
三组实验结果如图4A、4B和4C所示,在其他条件不变的情况下控制pH 4-9,温度20-50℃,时间0-720秒等条件下进行多组反应。最终确定pH为7.0、温度37℃、时间在720秒为最佳的实验条件。
因为乙酰胆碱酯酶能使乙酰胆碱分解失活,导致乙酰胆碱失去模拟酶活性,无法催化过氧化氢的分解,从而使的TMB溶液无法产生显色反应。
实施例1
一种生物传感器的制备方法,包括以下步骤:
将乙酰胆碱缓冲溶液和乙酰胆碱酯酶缓冲溶液混合在37℃培养10--15min,将混合溶液加入含过氧化氢的TMB溶液中,即可。
实施例2
一种生物传感器,采用实施例所述的方法制备得到。
实施例3
一种生物传感器检测乙酰胆碱酯酶的应用,具体检测方法为:
将乙酰胆碱缓冲溶液和分别与不同浓度的乙酰胆碱酯酶缓冲溶液混合,37℃培养15分钟,将混合溶液加入含过氧化氢的TMB溶液中,酰胆碱终浓度为10mM/L,乙酰胆碱酯酶终浓度分别为0.2mU/mL、0.4mU/mL、0.6mU/mL、0.8mU/mL、1.0mU/mL、1.2mU/mL和1.4mU/mL。利用紫外可见光谱仪器测出吸收波,构建吸光度与乙酰胆碱酯酶浓度的线性关系,在652nm处的吸光度做线性,得出了线性的标准曲线方程A=-0.10885C+0.1937,相关系数R2=0.99729,进而实现对乙酰胆碱酯酶的检测。结果如图6A、6B。
实施例4
一种检测有机磷农药的方法,具体检测方法为:
将乙酰胆碱酯酶缓冲溶液分别和不同浓度的的有机磷农药混合,在37℃反应10分钟,然后分别加入乙酰胆碱(0.01mM/L)中,最后所述混合液分别加入到含有过氧化氢的3,3',5,5'-四甲基联苯胺(TMB)溶液中发生显色反应,有机磷农药终浓度分别为10mU/uL、20mU/uL、30mU/uL、40mU/uL、50mU/uL、60mU/uL、70mU/uL、80mU/uL、90mU/uL、100mU/uL、110mU/uL、120mU/uL、130mU/uL、140mU/uL,乙酰胆碱酯酶终浓度为1.4mU/mL。通过紫外可见光谱仪器测出吸收波,然后在652nm处的吸光度做线性,最后得出了线性的标准曲线方程A=0.00189C+0.1679,相关系数R2=0.99323。
Claims (4)
1.一种生物传感器检测乙酰胆碱酯酶的应用,其特征在于,具体检测方法为:
将乙酰胆碱缓冲溶液和不同浓度的乙酰胆碱酯酶缓冲溶液混合培养,将混合溶液加入含过氧化氢的TMB溶液中,检测吸收波,构建吸光度与乙酰胆碱酯酶浓度的线性关系,进而实现对乙酰胆碱酯酶的检测;
乙酰胆碱终浓度为10mM/L,乙酰胆碱酯酶终浓度分别为0.2mU/mL、0.4mU/mL、0.6mU/mL、0.8mU/mL、1.0mU/mL、1.2mU/mL和1.4mU/mL。
2.根据权利要求1所述的生物传感器检测乙酰胆碱酯酶的应用,其特征在于,所述混合培养是指35℃-40℃培养10-15min。
3.根据权利要求1或2所述的生物传感器检测乙酰胆碱酯酶的应用,其特征在于,得到的线性的标准曲线方程A=-0.10885C+0.1937,相关系数的平方R2=0.99729。
4.一种生物传感器检测有机磷农药的方法,其特征在于,具体检测方法为:
将乙酰胆碱酯酶缓冲溶液分别与不同浓度的有机磷农药混合,得混合溶液,然后分别加入乙酰胆碱缓冲溶液中,最后将所得混合液分别加入到含有过氧化氢的TMB溶液中发生显色反应,检测吸收波,构建有机磷农药浓度与吸光度的线性关系,进而实现对有机磷农药的检测。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1508262A (zh) * | 2002-12-18 | 2004-06-30 | ҽѧ�о�Ժ | 乙酰胆碱酯酶分析 |
CN1731157A (zh) * | 2005-08-16 | 2006-02-08 | 中国检验检疫科学研究院 | 农药残毒快速检测用液体酶试剂及其制备方法与使用方法 |
CN102455295A (zh) * | 2010-10-28 | 2012-05-16 | 中国科学院上海生命科学研究院 | 用于有机磷和氨基甲酸酯类农药残留快速检测的显色方法 |
CN106399457A (zh) * | 2016-09-26 | 2017-02-15 | 南京大学 | 基于纳米模拟酶的可视化快速检测生物酶、蛋白质及其抑制剂的方法 |
CN106501424A (zh) * | 2016-11-10 | 2017-03-15 | 嘉兴职业技术学院 | 基于自动化前处理系统的农残检测方法 |
-
2017
- 2017-10-31 CN CN201711054416.1A patent/CN107817241B/zh active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1508262A (zh) * | 2002-12-18 | 2004-06-30 | ҽѧ�о�Ժ | 乙酰胆碱酯酶分析 |
CN1731157A (zh) * | 2005-08-16 | 2006-02-08 | 中国检验检疫科学研究院 | 农药残毒快速检测用液体酶试剂及其制备方法与使用方法 |
CN102455295A (zh) * | 2010-10-28 | 2012-05-16 | 中国科学院上海生命科学研究院 | 用于有机磷和氨基甲酸酯类农药残留快速检测的显色方法 |
CN106399457A (zh) * | 2016-09-26 | 2017-02-15 | 南京大学 | 基于纳米模拟酶的可视化快速检测生物酶、蛋白质及其抑制剂的方法 |
CN106501424A (zh) * | 2016-11-10 | 2017-03-15 | 嘉兴职业技术学院 | 基于自动化前处理系统的农残检测方法 |
Non-Patent Citations (2)
Title |
---|
Colorimetric determination of the activity of acetylcholinesterase and its inhibitors by exploiting the iodide-catalyzed oxidation of 3,3′,5,5′-tetramethylbenzidine by hydrogen peroxide;Pengjuan Ni et al.;《Microchim Acta》;20160930;第183卷(第9期);第2589-2595页 * |
Sihua Qian et al..Colorimetric Sensor Array for Detection and Identi fication of Organophosphorus and Carbamate Pesticides.《Analytical Chemistry》.2015,第87卷(第10期),第5395-5400页. * |
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