CN105973827A - 制备模拟抗体的分子印迹新方法及其细菌检测上的应用 - Google Patents
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Abstract
本发明所述的制备模拟抗体的分子印迹新方法及其细菌检测上的应用，是采用至少两种不同特性且具有不同功能基团的聚合物按比例共混制备分子印迹膜。本发明独特地将无交联的共混聚合物用于模拟抗体的分子印迹技术，通过聚合物不同功能基团与细菌表面之间模拟抗原‑抗体相互作用，增加印迹膜与细菌间的相互作用位点，从而提高印迹膜的特异性识别能力。本发明所述方法可以解决表面分子印迹技术特异性识别能力差、灵敏度低的问题，可提供一种特异性识别能力强、高灵敏度、成本低、简单易行的细菌检测方法。
Description
技术领域
本发明涉及一种基于界面诱导组装法制备高仿真模拟抗体的分子印迹新方法及其细菌检测上的应用。
背景技术
分子印迹技术（Molecular Imprinting Technique,
MIT）是人工合成与模板分子耦合的聚合物的一种新型实验制备技术，其基本思想源于人们对抗原-抗体以及酶-底物的专一性选择的认识，当模板分子与聚合物单体接触时形成多重作用点，通过聚合过程这种作用会被记忆下来，当把模板分子去除后，聚合物中就形成了与模板分子空间构型相匹配的具有多重作用点的空穴，这样的空穴将对模板分子及其类似物具有选择识别特性。通过MIT制备出的在空间结构和结合位点上与模板分子完全匹配的聚合物也被称为分子印迹聚合物（MIP）。MIT技术自德国Heinrich Heine大学Wuff和Sarhan首次报道人工合成分子印迹聚合物至今，已有将近40年的发展历史。从其发展过程来看，MIT在有机小分子领域的研究已相对成熟，已广泛应用于色谱分离、固相萃取、临床药物分析、化学仿生传感器等；在生物大分子领域的应用相对较少，大部分的研究机构都将重点都投入到生物大分子领域，该技术目前正在从有机小分子向生物大分子发展。而分子印迹聚合物的物理形式则正从三维块状分子印迹聚合物向二维薄膜分子印迹聚合物发展。虽然3D包埋法制备的印迹聚合物具有很好的识别选择性，但是模板分子分布于整个聚合物中，使得部分识别位点被包埋，造成空间位阻，导致聚合物吸附容量受到影响。2D表面印迹法则克服了3D包埋法的空间位阻问题，表面印迹法在薄膜表面形成印迹聚合物，识别位点暴露在载体的表面，使得模板分子能够自由进入或离开聚合物的识别位点。现有的技术多通过聚合物共混交联制备印迹膜，其高交联度易使MIP外部形态松散易碎，模板分子传质过程缓慢，吸附和脱附困难，无法有效的解决MIP特异性吸附效率的问题。同时，现有的表面印迹法的研究大都维持在空间结构相似的作用上，相互作用位点少且单一，导致其特异性识别能力差，影响了表面印迹法在检测领域的推广及应用。
发明内容
本发明的目的在于针对现有的表面分子印迹技术特异性识别能力差、灵敏度低的问题，提供一种制备具有特异性识别能力强、高灵敏度、成本低、简单易行的模拟抗体的分子印迹新方法，且在细菌检测上的应用效果优越。
本发明的技术方案是这样实现的：
本发明所述的制备模拟抗体的分子印迹新方法，其特点是：先将至少两种不同特性且具有不同功能基团的聚合物按一定比例共混成溶液，再将灭活的细菌溶液与该共混溶液混合，然后利用旋转涂膜法将混合溶液旋涂于多孔金膜上，并在常温下缓慢干燥成复合有细菌的共混膜，而在成膜的过程中，聚合物与细菌表面会发生诱导组装，不同功能基团与细菌表面之间形成模拟抗原-抗体的相互识别的位点，随后将复合有细菌的共混膜洗脱细菌模板，再在常温下缓慢干燥成分子印迹膜。
其中，所述不同功能基团为正负电荷和亲疏水基团。
所述共混为非交联共混。
所述共混膜的厚度小于细菌的直径。
本发明所述的模拟抗体的分子印迹在细菌检测上的应用，其特点是：将制备的分子印迹膜置于灭活的细菌样品溶液中吸附细菌，利用紫外分光光度计检测分子印迹膜吸附细菌前后吸收光谱峰值的移动变化，进而用于检测分子印迹膜对细菌特异性吸附能力。
本发明与现有技术相比，具有如下优点：
1.本发明利用多种的聚合物共混，通过分子间相互作用提高共混膜的力学性能，同时聚合物不交联，使得模板分子的传递过程更为顺畅，能很好的应用于印迹膜材料；
2. 由于不同特性的聚合物具有不同的功能基团（如正负电荷基团、亲疏水基团等），在与细菌的充分接触过程中，聚合物与细菌表面会发生诱导组装，聚合物的不同功能基团与细菌表面形成模拟抗原-抗体的相互识别位点，从而极大地增加了印迹膜与细菌间的相互作用位点，使得印迹膜的特异性识别能力大大提高；
因此，本发明采用的基于界面诱导组装法制备高仿真模拟抗体的分子印迹及其检测灭活细菌的应用具有广泛的推广应用价值。
下面结合附图对本发明作进一步的说明。
附图说明
图1为界面诱导组装原理图。
图2为高仿真模拟抗体分子印迹技术检测灭活细菌的过程图。
具体实施方式
本发明所述的制备模拟抗体的分子印迹新方法，先将至少两种不同特性且具有不同功能基团的聚合物按一定比例共混成溶液，再将灭活的细菌溶液与该共混溶液混合，然后利用旋转涂膜法将混合溶液旋涂于多孔金膜上，且旋涂于多孔金膜上的混合溶液是在常温下缓慢干燥成复合有细菌的共混膜，而在成膜的过程中，聚合物与细菌表面会发生诱导组装（如图1所示），不同功能基团与细菌表面之间形成模拟抗原-抗体的相互识别的位点，随后将复合有细菌的共混膜洗脱细菌模板，再在常温下缓慢干燥成分子印迹膜。其中，所述不同功能基团为正负电荷和亲疏水基团。所述共混为非交联共混。所述共混膜的厚度小于细菌的直径。
如图2所示，本发明所述的模拟抗体的分子印迹在细菌检测上的应用，是将制备的分子印迹膜置于灭活的细菌样品溶液中吸附细菌，利用紫外分光光度计检测分子印迹膜吸附细菌前后吸收光谱峰值的移动变化，进而用于检测分子印迹膜对细菌特异性吸附能力。
下面结合具体实施例对本发明作进一步的说明。
实施例1：
将聚乙烯醇（PVA）放置于水中，升温至80℃，恒温搅拌至其完全溶解；将明胶放置于水中，升温至40℃，恒温搅拌至其完全溶解；然后将制成的PVA溶液与明胶溶液按质量比为9:1的比例混合，随后再将灭活大肠杆菌溶液与PVA/明胶的共混溶液混合，利用旋转涂膜法将混合溶液旋涂在多孔金膜上，涂覆厚度为1μm，在常温下缓慢干燥成膜。将旋涂制备出的复合有大肠杆菌的明胶/聚乙烯醇膜（MIP膜），先在4℃环境下用溶菌酶（10mg/ml）预处理MIP膜2小时，将大肠杆菌细胞壁水解破坏，再用10%的Triton X处理MIP膜 80分钟，以除去对细胞壁和聚合物表面多糖之间的强相互作用，随后是用大量蒸馏水洗净MIP膜，最后将MIP膜在常温下干燥成膜。将旋涂在多孔金膜表面的MIP膜置于灭活大肠杆菌溶液中，25℃恒温振荡吸附细菌24h。利用紫外分光光度计检测该MIP膜吸附细菌前后的吸收光谱峰值的移动变化，对比现有的表面印迹膜吸附细菌前后的吸收光谱峰值的移动变化。
实施例2：
将琼脂糖放置于水中，升温至90℃以上，恒温搅拌至其完全溶解；将壳聚糖放置于水中，调节水中PH值至弱酸，恒温搅拌至其完全溶解；然后将制成的琼脂糖溶液与壳聚糖溶液按质量比为1:1的比例混合，随后再将灭活大肠杆菌溶液与琼脂糖/壳聚糖的共混溶液混合，利用旋转涂膜法将混合溶液旋涂在多孔金膜上，涂覆厚度为1μm，在常温下缓慢干燥成膜。将旋涂制备出的复合有大肠杆菌的琼脂糖/壳聚糖膜（MIP膜），先在4℃环境下用溶菌酶（10mg/ml）预处理MIP膜2小时，将大肠杆菌细胞壁水解破坏，再用10%的Triton X处理MIP膜 80分钟，以除去对细胞壁和聚合物表面多糖之间的强相互作用，随后是用大量蒸馏水洗净MIP膜，最后将MIP膜在常温下干燥成膜。将旋涂在多孔金膜表面的MIP膜置于灭活大肠杆菌溶液中，25℃恒温振荡吸附细菌24h。利用紫外分光光度计检测该MIP膜吸附细菌前后的吸收光谱峰值的移动变化，对比现有的表面印迹膜吸附细菌前后的吸收光谱峰值的移动变化。
综上，本研究是发明了一种避免共混高交联的影响，同时增加作用位点，高度仿真模拟抗原-抗体相互作用结合位点的细菌检测方法，提高了分子印迹技术的特异性识别能力和灵敏度，同时成本低且简单易行。
本发明是通过实施例来描述的，但并不对本发明构成限制，参照本发明的描述，所公开的实施例的其他变化，如对于本领域的专业人士是容易想到的，这样的变化应该属于本发明权利要求限定的范围之内。
Claims (5)
1.一种制备模拟抗体的分子印迹新方法，其特征在于：先将至少两种不同特性且具有不同功能基团的聚合物按一定比例共混成溶液，再将灭活的细菌溶液与该共混溶液混合，然后利用旋转涂膜法将混合溶液旋涂于多孔金膜上，并在常温下缓慢干燥成复合有细菌的共混膜，而在成膜的过程中，聚合物与细菌表面会发生诱导组装，不同功能基团与细菌表面之间形成模拟抗原-抗体的相互识别的位点，随后将复合有细菌的共混膜洗脱细菌模板，再在常温下缓慢干燥成分子印迹膜。
2.根据权利要求1所述的制备模拟抗体的分子印迹新方法，其特征在于：所述不同功能基团为正负电荷和亲疏水基团。
3.根据权利要求1所述的制备模拟抗体的分子印迹新方法，其特征在于：所述共混为非交联共混。
4.根据权利要求1所述的制备模拟抗体的分子印迹新方法，其特征在于：所述共混膜的厚度小于细菌的直径。
5.一种如上述任一权利要求所述分子印迹在细菌检测上的应用，其特征在于：将制备的分子印迹膜置于灭活的细菌样品溶液中吸附细菌，利用紫外分光光度计检测分子印迹膜吸附细菌前后吸收光谱峰值的移动变化，进而用于检测分子印迹膜对细菌特异性吸附能力。
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101775103A (zh) * | 2009-12-29 | 2010-07-14 | 湖北工业大学 | 一种蛋白质分子印迹薄膜的制备方法 |
| CN102105493A (zh) * | 2008-06-27 | 2011-06-22 | 哥伦比亚生物系统公司 | 用于检测微生物的分子印迹聚合物 |
| CN102514261A (zh) * | 2011-12-20 | 2012-06-27 | 中国科学院海洋研究所 | 一种微生物生物印迹薄膜及其制备方法 |
| EP2507278A1 (en) * | 2009-12-01 | 2012-10-10 | Cranfield University | Preparation of molecularly imprinted polymers |
| CN104945623A (zh) * | 2015-07-03 | 2015-09-30 | 华中科技大学 | 一种抗病毒分子印迹聚合物的制备方法 |
| CN105327684A (zh) * | 2015-12-03 | 2016-02-17 | 湖北出入境检验检疫局检验检疫技术中心 | 识别莫西菌素的磁性荧光分子印迹材料及其制备方法 |
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| EP2507278A1 (en) * | 2009-12-01 | 2012-10-10 | Cranfield University | Preparation of molecularly imprinted polymers |
| CN101775103A (zh) * | 2009-12-29 | 2010-07-14 | 湖北工业大学 | 一种蛋白质分子印迹薄膜的制备方法 |
| CN102514261A (zh) * | 2011-12-20 | 2012-06-27 | 中国科学院海洋研究所 | 一种微生物生物印迹薄膜及其制备方法 |
| CN104945623A (zh) * | 2015-07-03 | 2015-09-30 | 华中科技大学 | 一种抗病毒分子印迹聚合物的制备方法 |
| CN105327684A (zh) * | 2015-12-03 | 2016-02-17 | 湖北出入境检验检疫局检验检疫技术中心 | 识别莫西菌素的磁性荧光分子印迹材料及其制备方法 |
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