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Efficient protein immobilization on polyethersolfone electrospun nanofibrous membrane via covalent binding for biosensing applications

Mahmoudifard, M ; Sharif University of Technology | 2016

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  1. Type of Document: Article
  2. DOI: 10.1016/j.msec.2015.09.007
  3. Publisher: Elsevier Ltd , 2016
  4. Abstract:
  5. In this paper we introduce novel strategy for antibody immobilization using high surface area electrospun nanofibrous membrane based on ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling chemistry. To present the high performance of proposed biosensors, anti-staphylococcus enterotoxin B (anti-SEB) was used as a model to demonstrate the utility of our proposed system. Polymer solution of polyethersolfone was used to fabricate fine nanofibrous membrane. Moreover, industrial polyvinylidene fluoride membrane and conventional microtiter plate were also used to compare the efficiency of antibody immobilization. Scanning electron microscopy images were taken to study the morphology of the membranes. The surface activation of nanofibrous membrane was done with the help of O2 plasma. PES nanofibrous membrane with carboxyl functional groups for covalent attachment of antibodies were treated by EDC/NHS coupling agent. The quantity of antibody immobilization was measured by enzyme-linked immuno sorbent assay (ELISA) method. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) spectroscopy was performed to confirm the covalent immobilization of antibody on membrane. Atomic force microscopy, scanning electron microscopy and invert fluorescence microscopy were used to analyze the antibody distribution pattern on solid surfaces. Results show that oxygen plasma treatment effectively increased the amount of antibody immobilization through EDC/NHS coupling chemistry. It was found that the use of nanofibrous membrane causes the improved detection signal of ELISA based biosensors in comparison to the standard assay carried out in the 96-well microtiter plate. This method has the potential to improve the ELISA-based biosensor and we believe that this technique can be used in various biosensing methods
  6. Keywords:
  7. Antibody ; Immobilization ; Atomic force microscopy ; Biosensors ; Coupling agents ; Electron microscopy ; Electrospinning ; Enzyme immobilization ; Fluorescence microscopy ; Fourier transform infrared spectroscopy ; Membranes ; Nanofibers ; Plasma applications ; Radioactive waste vitrification ; Scanning electron microscopy ; 96-Well microtiter plate ; Attenuated total reflectance fourier transform infrared spectroscopies (ATR FTIR) ; Electrospun nanofibrous membranes ; Enzyme-linked immunosorbent assays ; Oxygen plasma treatments ; Polyvinylidene fluoride membranes ; Scanning electron microscopy image ; Antibodies ; Bacterium antibody ; Immobilized antibody ; Nanofiber ; Polyethersulfone ; Polymer ; Sulfone ; Animal ; Chemistry ; Devices ; Electrochemical analysis ; Genetic procedures ; Mouse ; Procedures ; Animals ; Antibodies, Bacterial ; Antibodies, Immobilized ; Biosensing Techniques ; Electrochemical Techniques ; Mice ; Nanofibers ; Polymers ; Protein Stability ; Sulfones
  8. Source: Materials Science and Engineering C ; Volume 58 , 2016 , Pages 586-594 ; 09284931 (ISSN)
  9. URL: http://www.sciencedirect.com/science/article/pii/S0928493115303362