Covalently derivatized NTA microarrays on porous silicon for multi-mode detection of His-tagged proteins  

作　　者：PEI Jia TANG YanChun XU Ning LU Wei XIAO ShouJun LIU JianNing 

机构地区：[1]State Key Laboratory of Coordination Chemistry,Nanjing National Laboratory for Microstructures School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China [2]State Key Laboratory of Bioelectronics Southeast University, Nanjing 210096, China [3]Institute of Molecular Medicine, Nanjing University, Nanjing 210093, China

出　　处：《Science China Chemistry》2011年第3期526-535,共10页中国科学（化学英文版）

基　　金：the financial support of the National Basic Research Program of China(2007CB925101);the National Natural Science Foundation of China(20721002&20827001);an open research fund of State Key Laboratory of Bioelectronics,Southeast University

摘　　要：Porous silicon(PSi)was applied as a supporting substrate for stepwise covalent derivatization of undecylenic acid, N-hydroxysuccinimidyl ester(NHS-ester)and nitrilotriacetic acid(NTA).By taking the advantages of porous silicon as a supporting matrix such as high surface area to volume ratio,infrared transparency,porous semiconductors for laser desorption/ionization mass spectroscopy,and low fluorescence background,a multi-mode detection biochip prototype can be realized. We prepared such a protein microarray by spotting NTA microarray dots on NHS-ester derivatized PSi,converting the rest of chip area into poly(ethylene glycol)background,loading NiII,and finally affinity-binding histidine-tagged(His-tagged)proteins.With the multi-mode analyses of infrared spectroscopy,X-ray photoelectron spectroscopy(XPS),atomic force microscopy(AFM),matrix-assisted laser desorption/ionization mass spectroscopy(MALDI-MS),and fluorescence scanning,two example proteins,His-tagged thioredoxin-urodilatin and His-tagged aprotinin,were well qualified and quantified.Porous silicon（PSi）was applied as a supporting substrate for stepwise covalent derivatization of undecylenic acid, N-hydroxysuccinimidyl ester（NHS-ester）and nitrilotriacetic acid（NTA）.By taking the advantages of porous silicon as a supporting matrix such as high surface area to volume ratio,infrared transparency,porous semiconductors for laser desorption/ionization mass spectroscopy,and low fluorescence background,a multi-mode detection biochip prototype can be realized. We prepared such a protein microarray by spotting NTA microarray dots on NHS-ester derivatized PSi,converting the rest of chip area into poly（ethylene glycol）background,loading NiII,and finally affinity-binding histidine-tagged（His-tagged）proteins.With the multi-mode analyses of infrared spectroscopy,X-ray photoelectron spectroscopy（XPS）,atomic force microscopy（AFM）,matrix-assisted laser desorption/ionization mass spectroscopy（MALDI-MS）,and fluorescence scanning,two example proteins,His-tagged thioredoxin-urodilatin and His-tagged aprotinin,were well qualified and quantified.

关 键 词：porous silicon MICROARRAY MULTI-MODE NTA His-tagged protein 

分 类 号：O614.21[理学—无机化学] Q51[理学—化学]