CN106018851B - 一种基于片状黑磷的类固醇免疫传感器的制备方法及应用 - Google Patents
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用 Download PDFInfo
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Abstract
本发明涉及一种基于片状黑磷的类固醇免疫传感器的制备方法及应用,属于新型生物传感检测技术领域。片状黑磷具有类似石墨烯性质的结构,具有良好的导电性。同时离子液体作为片状黑磷的分散剂,将其应用在免疫传感器的基底。同时,利用多孔PtAu合金膜将底层材料和抗体牢牢地结合起来,增强传感器的稳定性。利用底液铁氰化钾为信号,实现了对类固醇的超灵敏检测。
Description
技术领域
本发明涉及一种基于片状黑磷的类固醇免疫传感器的制备方法及应用,采用黑磷@离子液体复合材料作为检测基底材料,增强导电性的同时,又能增加信号分子在传感表面的固载量上,通过多孔PtAu合金膜的修饰,制备一种检测类固醇的电化学免疫传感器,属于新型功能材料与生物传感检测技术领域。
技术背景
类固醇是广泛分布于生物界的一大类环戊稠全氢化菲衍生物的总称。就雌二醇而言,它可以补充雌激素不足,常用于治疗女性性腺功能不良、双侧卵巢切除术后、更年期综合征如潮热、出汗和精神、神经症状等。但是会有不良反应,如腹部绞痛或胀气,恶心,踝及足水肿等。
目前对于类固醇的检测方法不是很多,主要以放射性免疫分析,气相色谱质谱,液相色谱质谱为主。但是这些方法需要大型设备及专业仪器,需要专业培训的技术人员进行操作。而免疫传感器是将免疫学方法与分析化学方法相结合的一种生物传感器,通过抗原与抗体之间的特异性结合,构建了操作简便、结构简单、选择性好、灵敏度高的免疫传感器。本发明制备了一种基于片状黑磷的类固醇免疫传感器。
本发明采用黑磷与离子液体作用形成的复合材料为基底材料,实现了信号分子的高通量负载。本发明还采用了多孔PtAu合金膜构建免疫传感器,既保证了稳定性,又增加了类固醇抗体的负载,实现了类固醇的灵敏检测。该方法具有成本低、特异性好、检测快速等优点,而且制备过程较为简单,有效克服了目前类固醇检测方法的不足。
发明内容
本发明的目的之一是基于黑磷@离子液体P@IL复合材料为基底材料,构建了一种无酶,超灵敏的电化学免疫传感器。
本发明的目的之二是通过多孔PtAu合金膜的使用,实现了对类固醇的超灵敏检测。
本发明的技术方案如下
1. 一种基于片状黑磷的类固醇免疫传感器的制备方法及应用
(1)将直径3 mm的玻碳电极用Al2O3抛光粉打磨,超纯水洗净;将1 ~ 10 µL、1 ~ 10mg/mL黑磷@离子液体P@IL复合材料滴加到电极表面,超纯水冲洗,室温下晾干成膜;
(2)将投影面积为5 ~ 10 mm2多孔PtAu合金膜覆盖在电极表面,超纯水冲洗;
(3)滴加1 ~ 10 µL类固醇抗体标准溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(4) 滴加1 ~ 10 µL质量分数为0.5% ~ 1.5%的牛血清白蛋白BSA溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(5)滴加1 ~ 10 µL、0.005 ~ 20 ng/mL的一系列不同浓度的类固醇抗原标准溶液至电极表面,超纯水冲洗,4℃冰箱中晾干。
2. 黑磷@离子液体P@IL复合材料的制备
取0.2 ~ 2.0 mg的片状黑磷加入到0.5 ~ 20 mL的离子液体1-丁基-3-甲基咪唑四氟硼酸盐BMIM∙BF4中,振荡12 h,得到复合材料黑磷@离子液体P@IL。
3. 多孔PtAu合金膜的制备
室温下,将投影面积为5 ~ 30 mm2的PtAuAg合金膜漂浮在16 mol/L的硝酸液面上,腐蚀10 ~ 100 min,将腐蚀后的合金膜用超纯水洗至中性,即得到多孔PtAu合金膜。
4. 一种基于片状黑磷的类固醇免疫传感器的制备方法及应用,所述的类固醇选自下列之一:雌酮,雌二醇,炔雌醇。
本发明有益成果
(1)黑磷的使用,能够很好的促进电极表面的电子传递,同时实现了对传感器底层比表面积成功放大,达到高通量负载。
(2)黑磷@离子液体P@IL复合材料的使用,使得生物分子的负载量大大提升,扩展了传感器的线性范围,降低了检出限。
(3)多孔PtAu合金膜的使用,增加了传感器表面的电子传输能力,增强了传感器的稳定性,实现了类固醇超灵敏稳定检测。
具体实施方式
实施例1
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用
(1)将直径3 mm的玻碳电极用Al2O3抛光粉打磨,超纯水洗净;将1 µL、1 mg/mL黑磷@离子液体P@IL复合材料滴加到电极表面,超纯水冲洗,室温下晾干成膜;
(2)将投影面积为5 mm2多孔PtAu合金膜覆盖在电极表面,超纯水冲洗;
(3)滴加1 µL类固醇抗体标准溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(4) 滴加1 µL质量分数为0.5%的牛血清白蛋白BSA溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(5)滴加1 µL、0.005 ~ 20 ng/mL的一系列不同浓度的类固醇抗原标准溶液至电极表面,超纯水冲洗,4℃冰箱中晾干。
实施例2
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用
(1)将直径3 mm的玻碳电极用Al2O3抛光粉打磨,超纯水洗净;将4 µL、4 mg/mL黑磷@离子液体P@IL复合材料滴加到电极表面,超纯水冲洗,室温下晾干成膜;
(2)将投影面积为7 mm2多孔PtAu合金膜覆盖在电极表面,超纯水冲洗;
(3)滴加5 µL类固醇抗体标准溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(4) 滴加5 µL质量分数为1.0%的牛血清白蛋白BSA溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(5)滴加5 µL、0.005 ~ 20 ng/mL的一系列不同浓度的类固醇抗原标准溶液至电极表面,超纯水冲洗,4℃冰箱中晾干。
实施例3
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用
(1)将直径3 mm的玻碳电极用Al2O3抛光粉打磨,超纯水洗净;将10 µL、10 mg/mL黑磷@离子液体P@IL复合材料滴加到电极表面,超纯水冲洗,室温下晾干成膜;
(2)将投影面积为10 mm2多孔PtAu合金膜覆盖在电极表面,超纯水冲洗;
(3) 滴加10 µL类固醇抗体标准溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(4) 滴加10 µL质量分数为1.5%的牛血清白蛋白BSA溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(5)滴加10 µL、0.005 ~ 20 ng/mL的一系列不同浓度的类固醇抗原标准溶液至电极表面,超纯水冲洗,4℃冰箱中晾干。
实施例4
黑磷@离子液体P@IL复合材料的制备
取0.2 mg的片状黑磷加入到0.5 mL的离子液体1-丁基-3-甲基咪唑四氟硼酸盐BMIM∙BF4中,振荡12 h,得到复合材料黑磷@离子液体P@IL。
实施例5
黑磷@离子液体P@IL复合材料的制备
取1.0 mg的片状黑磷加入到10 mL的离子液体1-丁基-3-甲基咪唑四氟硼酸盐BMIM∙BF4中,振荡12 h,得到复合材料黑磷@离子液体P@IL。
实施例6
黑磷@离子液体P@IL复合材料的制备
取2.0 mg的片状黑磷加入到20 mL的离子液体1-丁基-3-甲基咪唑四氟硼酸盐BMIM∙BF4中,振荡12 h,得到复合材料黑磷@离子液体P@IL。
实施例7
多孔PtAu合金膜的制备
室温下,将投影面积为5 mm2的PtAuAg合金膜漂浮在16 mol/L的硝酸液面上,腐蚀10 min,将腐蚀后的合金膜用超纯水洗至中性,即得到多孔PtAu合金膜。
实施例8
多孔PtAu合金膜的制备
室温下,将投影面积为10 mm2的PtAuAg合金膜漂浮在16 mol/L的硝酸液面上,腐蚀50 min,将腐蚀后的合金膜用超纯水洗至中性,即得到多孔PtAu合金膜。
实施例9
多孔PtAu合金膜的制备
室温下,将投影面积为30 mm2的PtAuAg合金膜漂浮在16 mol/L的硝酸液面上,腐蚀100 min,将腐蚀后的合金膜用超纯水洗至中性,即得到多孔PtAu合金膜。
实施例10
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用,所述的类固醇为雌酮。
实施例11
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用,所述的类固醇为雌二醇。
实施例12
一种基于片状黑磷的类固醇免疫传感器的制备方法及应用,所述的类固醇为炔雌醇。
Claims (4)
1.一种基于片状黑磷的类固醇免疫传感器的制备方法,其特征在于,包括以下步骤:
(1)将直径3 mm的玻碳电极用Al2O3抛光粉打磨,超纯水洗净;将1 ~ 10 µL、1 ~ 10 mg/mL黑磷@离子液体P@IL复合材料滴加到电极表面,超纯水冲洗,室温下晾干成膜;
(2)将投影面积为5 ~ 10 mm2多孔PtAu合金膜覆盖在电极表面,超纯水冲洗;
(3)滴加1 ~ 10 µL类固醇抗体标准溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(4) 滴加1 ~ 10 µL质量分数为0.5% ~ 1.5%的牛血清白蛋白BSA溶液于电极表面,超纯水冲洗,4℃冰箱中晾干;
(5)滴加1 ~ 10 µL、0.005 ~ 20 ng/mL的一系列不同浓度的类固醇抗原标准溶液至电极表面,超纯水冲洗,4℃冰箱中晾干。
2.如权利要求1所述的一种基于片状黑磷的类固醇免疫传感器的制备方法,其特征在于:所述的黑磷@离子液体P@IL复合材料的制备方法,包括以下步骤:
取0.2 ~ 2.0 mg的片状黑磷加入到0.5 ~ 20 mL的离子液体1-丁基-3-甲基咪唑四氟硼酸盐BMIM∙BF4中,振荡12 h,得到复合材料黑磷@离子液体P@IL。
3.如权利要求1所述的一种基于片状黑磷的类固醇免疫传感器的制备方法,其特征在于:所述的多孔PtAu合金膜的制备方法,包括以下步骤:
室温下,将投影面积为5 ~ 10 mm2的PtAuAg合金膜漂浮在16 mol/L的硝酸液面上,腐蚀10 ~ 100 min,将腐蚀后的合金膜用超纯水洗至中性,即得到多孔PtAu合金膜。
4.如权利要求1所述的一种基于片状黑磷的类固醇免疫传感器的制备方法,所述的类固醇选自下列之一:雌酮,雌二醇,炔雌醇。
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WO2020113362A1 (zh) * | 2018-12-03 | 2020-06-11 | 中国水产科学研究院 | 一种酪氨酸酶修饰的黑磷及其在生物传感器中的应用 |
CN110687182A (zh) * | 2019-10-30 | 2020-01-14 | 云南大学 | 一种检测前列腺特异性抗原的电化学免疫传感器的制备方法 |
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CN111678962B (zh) * | 2020-06-17 | 2022-07-01 | 山东理工大学 | 一种黄曲霉毒素电化学传感器的制备方法及应用 |
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