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Preparation of protein imprinted materials by hierarchical imprinting techniques and application in selective depletion of albumin from human serum

机译:通过分层压印技术制备蛋白质印迹材料及其在人血清中白蛋白的选择性耗尽中的应用

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Hierarchical imprinting was developed to prepare the protein imprinted materials, as the artificial antibody, for the selective depletion of HSA from the human serum proteome. Porcine serum albumin (PSA) was employed as the dummy template for the fabrication of the recognition sites. To demonstrate the advantages of the hierarchical imprinting, molecularly imprinted polymers prepared by hierarchical imprinting technique (h-MIPs) were compared with those obtained by bulk imprinting (b-MIPs), in terms of the binding capacity, adsorption kinetics, selectivity and synthesis reproducibility. The binding capacity of h-MIPs could reach 12?mg g?1. And saturation binding could be reached in less than 20?min for the h-MIPs. In the protein mixture, h-MIPs exhibit excellent selectivity for PSA, with imprinting factors as about 3.6, much higher than those for non-template proteins. For the proteomic application, the identified protein group number in serum treated by h-MIPs was increased to 422, which is 21% higher than that obtained from the original serum, meanwhile the identified protein group number for the Albumin Removal kit was only 376. The results demonstrate that protein imprinted polymers prepared by hierarchical imprinting technique, might become the artificial antibodies for the selective depletion of high abundance proteins in proteome study.
机译:开发了分层印刷,以制备蛋白质印迹材料,作为人工抗体,用于从人血清蛋白质中选择性耗尽HSA。猪血清白蛋白(PSA)用作制备识别位点的伪模板。为了证明等级印迹的优点,将通过分层压印技术(H-MIPS)制备的分子印迹聚合物与通过块状压印(B-MIPS)获得的那些,就结合能力,吸附动力学,选择性和合成再现性而获得。 H-MIPS的绑定容量可以达到12?Mg G ?1 。对于H-MIPS,可以在少于20℃的少于20·min的情况下达到饱和结合。在蛋白质混合物中,H-MIPS对PSA具有优异的选择性,具有约3.6的印迹因子,远高于非模板蛋白的因素。对于蛋白质组学应用,通过H-MIPS处理的血清中鉴定的蛋白质组数升至422,其比从原始血清中获得的21%,同时白蛋白去除试剂盒的鉴定的蛋白质组数仅为376。结果表明,通过分级压印技术制备的蛋白质印迹聚合物,可能成为蛋白质组研究中选择性脱落的人工抗体。

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