Lysozyme-imprinted polymer synthesized using UV free-radical polymerization.
Lysozyme-imprinted polymer synthesized using UV free-radical polymerization.
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DOI：
10.1016/j.talanta.2010.08.055
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发表时间：
2010-11
期刊：
影响因子：
6.1
通讯作者：
Shuang Yu;Shuang Yu;Aiqin Luo;D. Biswal;J. Z. Hilt;D. Puleo
中科院分区：
文献类型：
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作者：
Shuang Yu;Shuang Yu;Aiqin Luo;D. Biswal;J. Z. Hilt;D. Puleo
分子印迹是一种制备能够选择性识别模板分子的高分子材料（分子印迹聚合物或MIP）的方法。小分子的分子印迹已被广泛研究。然而，不太常见的是生物大分子的印迹，包括蛋白质，其中溶菌酶是食品，制药和诊断科学中的重要分子。本研究采用两步法制备了溶菌酶分子印迹聚合物。首先，溶菌酶，PEG600DMA，和甲基丙烯酸分别作为模板分子，交联单体，和功能单体，在紫外自由基聚合过程中合成的聚合物凝胶。其次，通过酶消化去除溶菌酶。在不添加溶菌酶的条件下合成了非印迹聚合物。为了评价MIP的优先结合能力，将溶菌酶、RNA酶A或溶菌酶和RNA酶A的50：50混合物加入MIP和NIP中，然后通过消化释放。结果发现，当更多的溶菌酶加入到反应混合物中时，从聚合物中释放的蛋白质的量增加，反映了更多的潜在结合位点。MIP与溶菌酶和RNase A的竞争结合混合物的测试显示MIP优先结合更大量的溶菌酶，高达RNase A的20倍。NIP仅结合少量的两种蛋白质，并且没有显示出对结合溶菌酶或RNA酶A的偏好。实验结果表明，利用紫外自由基聚合法成功地将溶菌酶分子印迹到分子印迹聚合物上，且所制备的分子印迹聚合物能够优先结合其模板蛋白。
Molecular imprinting is a method to fabricate a polymeric material (molecularly imprinted polymer or MIP) capable of selectively recognizing template molecules. Molecular imprinting of small molecules has been studied widely. Less common, however, is the imprinting of biological macromolecules, including proteins, among which lysozyme is an important molecule in the food, pharmaceutical, and diagnostic sciences. In this study, lysozyme MIP was fabricated in two steps. First, lysozyme, PEG600DMA, and methacrylic acid were used as the template molecule, cross-linking monomer, and the functional monomer, respectively, in a UV free-radical polymerization process to synthesize a polymeric gel. Second, lysozyme was removed by enzymatic digestion. Non-imprinted polymer (NIP) was synthesized without lysozyme addition. To evaluate the preferential binding capability of MIP, lysozyme, RNase A, or a 50:50 mixture of lysozyme and RNase A was added to MIP and NIP and then released by digestion. It was found that when more lysozyme was added to the reaction mixture, the quantity of protein released from the polymer increased, reflecting more potential binding sites. Tests of MIP with a competitive binding mixture of lysozyme and RNase A showed the MIP preferentially bound a greater amount of lysozyme, up to 20 times more than RNase A. NIP bound only small amounts of both proteins and did not show a preference for binding either lysozyme or RNase A. These results demonstrate that lysozyme was successfully imprinted into the MIP by UV free-radical polymerization, and the fabricated MIP was able to preferentially bind its template protein.