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Thiol Modified Chitosan Self-Assembled Monolayer Platform for Nucleic Acid Biosensor

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Abstract

A self-assembled monolayer (SAM) of thiol modified chitosan (SH-CHIT), with thioglycolic acid (TGA) as a modifier to bestow thiol groups, has been prepared onto gold (Au)-coated glass plates for fabrication of the nucleic acid biosensor. The chemical modification of CHIT via TGA has been evidenced by Fourier transform infrared spectroscopy (FT-IR) studies, and the biocompatibility studies reveal that CHIT retains its biocompatible nature after chemical modification. The electrochemical studies conducted onto SH-CHIT/Au electrode reveal that thiol modification in CHIT amino end enhances the electrochemical behavior indicating that it may be attributed to delocalization of electrons in CHIT skeleton that participates in the resonance process. The carboxyl group modified end of DNA probe has been immobilized onto SH-CHIT/Au electrode using N-ethyl-N′-(3-dimethylaminopropyl)carbodimide (EDC) and N-hydroxysuccinimide (NHS) chemistry for detection of complementary, one-base mismatch and non-complementary sequence using electrochemical and optical studies for Mycobacterium tuberculosis detection. It has been found that DNA-SH-CHIT/Au bioelectrode can specifically detect 0.01 μM of target DNA concentration with sensitivity of 1.69 × 10−6 A μM−1.

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Acknowledgments

We thank Prof. R.C. Budhani, Director, National Physical Laboratory, India for the facilities. Maumita Das Mukherjee is thankful to Amity University, Noida for allowing her to perform the experiments. The financial support received at NPL, from Department of Science and Technology, India under the projects GAP 081132 and GAP 080232 is greatly acknowledged.

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Authors and Affiliations

  1. Amity Institute of Applied Sciences, Amity University, Noida, Uttar Pradesh, India

    Maumita Das Mukherjee

  2. Special Centre for Nanoscience, Jawaharlal Nehru University, New Delhi, India

    Pratima R. Solanki

  3. Department of Science and Technology Centre on Biomolecular Electronics, Biomedical Instrumentation Section, Materials Physics and Engineering Division, CSIR- National Physical Laboratory, New Delhi, 110012, India

    Gajjala Sumana & Bansi D. Malhotra

  4. Department of Physical Electronics, Tokyo Institute of Technology, 2-12-1 O-okayama Meguro-ku, Tokyo, 152-8552, Japan

    Takaaki Manaka & Mitsumasa Iwamoto

  5. Department of Biotechnology, Delhi Technological University, Main Bawana Road, Shahbad Daulatpur, Delhi, 110042, India

    Bansi D. Malhotra

Authors
  1. Maumita Das Mukherjee
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  2. Pratima R. Solanki
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  3. Gajjala Sumana
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  4. Takaaki Manaka
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  5. Mitsumasa Iwamoto
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  6. Bansi D. Malhotra
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Correspondence to Bansi D. Malhotra.

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Mukherjee, M.D., Solanki, P.R., Sumana, G. et al. Thiol Modified Chitosan Self-Assembled Monolayer Platform for Nucleic Acid Biosensor. Appl Biochem Biotechnol 174, 1201–1213 (2014). https://doi.org/10.1007/s12010-014-1177-7

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  • DOI: https://doi.org/10.1007/s12010-014-1177-7

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