典型文献
A preparation strategy for protein-oriented immobilized silica magnetic beads with Spy chemistry for ligand fishing
文献摘要：
Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1-6)for 2 h,under 20％ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding.
文献关键词：
中图分类号：
作者姓名：
Yu Yi;Jianming Hu;Shenwei Ding;Jianfeng Mei;Xudong Wang;Yanlu Zhang;Jianshu Chen;Guoqing Ying
作者机构：
Institute of Biopharmaceuticals,College of Pharmaceutical Science,Zhejiang University of Technology,Hangzhou,310014,China
文献出处：
引用格式：
[1]Yu Yi;Jianming Hu;Shenwei Ding;Jianfeng Mei;Xudong Wang;Yanlu Zhang;Jianshu Chen;Guoqing Ying-.A preparation strategy for protein-oriented immobilized silica magnetic beads with Spy chemistry for ligand fishing)[J].药物分析学报（英文）,2022(03):415-423
A类：
hydroxysulfosuccinimide,spontaneity,isopeptide,unbreakable
B类：
preparation,strategy,oriented,immobilized,silica,magnetic,beads,chemistry,ligand,fishing,Due,complexity,bioactive,ingredients,biological,samples,screening,target,proteins,process,Herein,feasible,directing,immobilization,SpyTag,SpyCatcher,ST,SC,mediated,anchoring,presented,Carboxyl,groups,surface,coated,SMBs,were,coupled,using,dimethylaminopropyl,carbodiimide,hydrochloride,method,named,green,fluorescent,GFP,capturing,model,was,labeled,anchored,orientation,onto,directly,from,relevant,cell,lysates,via,self,ligation,characteristics,studied,electron,microscopy,energy,dispersive,spectroscopy,Fourier,transform,infrared,site,specificity,this,unique,reaction,confirmed,electrophoresis,fluorescence,analyses,Although,alkaline,stability,ligated,not,ideal,formed,bond,under,acidic,conditions,glycine,HCl,buffer,ethanol,solution,within,days,most,temperatures,We,therefore,simple,universal,diverse,functionalized,prompting,its,usage,drug,finding
AB值：
0.591586
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