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

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Abstract

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.

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Authors and Affiliations

  1. State Key Laboratory of Coordination Chemistry; Nanjing National Laboratory for Microstructures; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210093, China

    Jia Pei, Ning Xu & ShouJun Xiao

  2. State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China

    ShouJun Xiao

  3. Institute of Molecular Medicine, Nanjing University, Nanjing, 210093, China

    YanChun Tang, Wei Lu & JianNing Liu

Authors
  1. Jia Pei
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  2. YanChun Tang
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  3. Ning Xu
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  4. Wei Lu
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  5. ShouJun Xiao
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  6. JianNing Liu
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Correspondence to ShouJun Xiao or JianNing Liu.

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Pei, J., Tang, Y., Xu, N. et al. Covalently derivatized NTA microarrays on porous silicon for multi-mode detection of His-tagged proteins. Sci. China Chem. 54, 526–535 (2011). https://doi.org/10.1007/s11426-010-4128-3

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  • DOI: https://doi.org/10.1007/s11426-010-4128-3

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