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Published on: September 30, 2019
Screening of phage-displayed human liver cDNA library against doxorubicin with drug-immobilized monolithic
Xiaoming Yu1, Peng Zhao, Lihua Zhang
1The Medical School, Southeast University, Nanjing, 210009, China; Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.
Abstract:
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.
