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首页> 外文期刊>Biomedical Chromatography: An International Journal Devoted to Research in Chromatographic Methodologies and Their Applications in the Biosciences >Screening of phage-displayed human liver cDNA library against doxorubicin with drug-immobilized monolithic polyacrylamide cryogel
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Screening of phage-displayed human liver cDNA library against doxorubicin with drug-immobilized monolithic polyacrylamide cryogel

机译:药物固定的整体聚丙烯酰胺冷冻凝胶筛选针对阿霉素的噬菌体展示的人肝cDNA文库

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摘要

Monolithic polyacrylamide cryogel was prepared and utilized as a new matrix for drug immobilization to screen against phage-displayed human liver cDNA library. The macropores and hydrophilic nature of the cryogel made it possible for phage particles to pass unhindered. Doxorubicin, an anticancer drug, was covalently bonded to the monolithic cryogel by the glutaraldehyde method, and after five rounds of affinity selection performed in an SPE cartridge, phage clones that displayed Homo sapiens methyl CpG binding protein 2 (MeCP2) were selectively enriched. The interaction between doxorubicin and MeCP2 displayed phages was further validated by studying the retention of doxorubicin on MeCP2 phage-coupled cryogel. These results demonstrate that drug-coupled polyacrylamide cryogel might be a promising kind of matrix for screening target proteins against phage-displayed library.
机译:制备了整体聚丙烯酰胺冷冻凝胶,并将其用作药物固定的新基质,以针对噬菌体展示的人肝cDNA文库进行筛选。冷冻凝胶的大孔和亲水特性使噬菌体颗粒可以不受阻碍地通过。通过戊二醛法将抗癌药阿霉素共价键合至整体式冷冻凝胶,并在SPE药盒中进行了五轮亲和力选择后,选择性富集了展示智人甲基CpG结合蛋白2(MeCP2)的噬菌体克隆。阿霉素和MeCP2展示的噬菌体之间的相互作用通过研究阿霉素在MeCP2噬菌体偶联的冷冻凝胶上的保留来进一步验证。这些结果表明,药物偶联的聚丙烯酰胺冷冻凝胶可能是一种有前途的用于筛选针对噬菌体展示文库的靶蛋白的基质。

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