Construction Of Anti Omp Immobilized Nanoporous Membrane Based Electrochemical Biosensor For The Detection Of E. Coli

Authors

  • Manjul Mungali1*, Veena Pandey, Mohommad K. P. Singh Department of Biotechnology, Kumaun University, Bhimtal Campus, Bhimtal, India Defense Institute of Bioenergy Research (DIBER), DRDO, Gaura Parao, Haldwani, India Bionanotechnology & Nanobiosensor Research Laboratory, Biophysics Unit, CBSH, G.B. Pant University of Agriculture & Technology, Pantnagar-263145, India.

Keywords:

E. coli, electrochemical biosensor, impedance, nanoporous membrane, FTIR

Abstract

The present work was to develop an electrochemical biosensor for the rapid detection of E. coli (DH5α) OmpA antigen. Here, we utilize carbodiimide based strategies to crosslink antibodies on amine-functionalized platforms for diagnostic applications. The main objective of this study was to optimize the surface of nanoporous polycarbonate trek etched membrane (PCTE) to provide efficient binding of anti- OmpA (mouse anti Escherichia coli 03 (NTCC 9003) antibody and to exploit sensing surface for the detection of OmpA antigen (Escherichia coli DH5 alpha (1652)), as an alternative diagnostic test. The PCTE membrane was characterized by infrared spectroscopy. The inferred spectra shows specific peak of functionalization of PCTE membrane by carbodiimide (EDC-NHS) by converting terminal carboxylic group to an active NHS ester to bind antibodies ( Ab) (mouse anti Escherichia coli 03 (NTCC 9003)) at 1772 cm-1 (C=O vibrations), 1406 cm-1  and 1079 cm-1 (C-OH vibrations), and 766 cm-1 (OCN vibrations); while peaks on 3387 cm-1(N-H stretch), 1639 cm-1 (C=O) and 1567 cm-1 (NH2 wag) exhibits specific Ab-Ag (OmpA (Escherichia coli DH5 alpha (1652)) interaction.  We used a specially designed electrochemical biosensor setup for impedance measurement to monitor Ab-Ag interaction. The limit of detection of constructed electrochemical biosensor was 10ng/ml.

References

Demarco D.R. and D.V. Lim. 2002. “Detection of Escherichia coli O157:H7 in 10-and 25- gram ground beef samples with an evanescent-wave biosensor with silica and polystyrene waveguides.” J. Food Protect. 65:596-602.

Deneer, H.G. and A.A. Potter. 1989. Iron-repressible outer-membrane proteins of Pasteurella. hemolytica. J. Gen. Microbiol., 135: 435-443.

Feng Yang, Xiaodong Xiao, Zhongyu Zhu, Emily Streaker, Mitchell Ho Ira Pastan, and Dimiter S. Dimitrov. 2009. Mol Cancer Ther 8(5):1113–8.

Geng T., J. Uknalis, S.-T. Tu and A.K. Bhunia 2006. Fiber-Optic Biosensor Employing Alexa-Fluor Conjugated Antibody for Detection of Escherichia coli O157:H7 from Ground Beef in Four Hours. Sensors 6:796-807.

Jung-Chih Chen, S. Sadhasivam, Feng-Huei Lin, Yi-Yuan Yang, Chi-Hsin Lee. 2011. A simple and sensitive detection of ompa protein from escherichia coli by quartz crystal microbalance. IPCBEE vol.2.

Kong Jilie and YU. Shaoning. 2007. Fourier transform infrared spectroscopic analysis of protein secondary structures. Acta Biochimica et Biophysica Sinica. 39(8): 549–559.

Lin Mouhong, Yingju Liu, Zhuohong Yang, Yibin Huang, Zihong Sun, Yuan He, Chunlin. 2012. Construction of sensitive amperometric immunosensor based on poly(amidoamine) dendrimer and one-step ionic-liquid-assisted graphene/chitosan platform for benzo[a]pyrene detection. Int. J. Electrochem. Sci., 7 965 – 978.

Liu Guozhen, Jingquan Liu, Thomas P. Davis, J. Justin Gooding. 2011. Electrochemical impedance immunosensor based on gold nanoparticles and aryl diazonium salt functionalized gold electrodes for the detection of antibody. Biosensors and Bioelectronics 26 3660–3665.

Mura S., G. Greppi, M.L. Marongiu, P.P. Roggero, A.P. Ravindranath, L.J. Mauer, N. Schibeci, N. Perria, M. Piccinini, P. Innocenzi and J. Irudayaraj. 2009. FTIR nanobiosensors for Escherichia coli detection. Beilstein J. Nanotechnol. 3:485–492. doi:10.3762/bjnano.3.55.

Park S. and R. S. Ruoff. 2009. Nat. Nanotechnol., 4, 217–224.

Sharma J., M. Sharma and P. Ray. 2010. Detection of TEM & SHV genes in Escherichia coli & Klebsiella pneumoniae isolates in a tertiary care hospital from India. Ind. J. Med. Res. 132:332-336.

Singh K.P., M.K. Choudhary, I. Chianella and P. Singh. 2013a. Development of non-labeled QCM biosensor for the detection of β-Galactosidase: A comparative study of gold and polystyrene nanoparticles. Adv. Nanoparticles 2:182-190.

Singh K.P., R.K. Prajapat, S. Ahlawat, M. Mungali, S. Kumar. 2013b. Use of isoproturon imprinted polymer membranes as a selective recognition platform in a resistance based electrochemical sensor. Open J. Appl. Biosensor 2: 20-28.

Downloads

Published

2015-03-31

Issue

Vol. 4 No. 4 (2015): MARCH 2015

Section

Articles

License

Copyright Notice

Submission of an article implies that the work described has not been published previously (except in the form of an abstract or as part of a published lecture or academic thesis), that it is not under consideration for publication elsewhere, that its publication is approved by all authors and tacitly or explicitly by the responsible authorities where the work was carried out, and that, if accepted, will not be published elsewhere in the same form, in English or in any other language, without the written consent of the Publisher. The Editors reserve the right to edit or otherwise alter all contributions, but authors will receive proofs for approval before publication.

Copyrights for articles published in World Scholars journals are retained by the authors, with first publication rights granted to the journal. The journal/publisher is not responsible for subsequent uses of the work. It is the author's responsibility to bring an infringement action if so desired by the author.