title: Efficient protein immobilization on polyethersolfone electrospun nanofibrous membrane via covalent binding for biosensing applications.
abstract: 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.
authors.affiliation: Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran, Iran.
authors.forename: Matin
authors.initials: M
authors.lastname: Mahmoudifard
authors.affiliation: Stem Cell Biology Department, Stem Cell Technology Research Center, Tehran, Iran; Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Sara
authors.initials: S
authors.lastname: Soudi
authors.affiliation: Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Nanotechnology and Tissue Engineering Department, Stem Cell Technology Research Center, Tehran, Iran; School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Simzar
authors.initials: S
authors.lastname: Hosseinzadeh
authors.affiliation: Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran, Iran; Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Elaheh
authors.initials: E
authors.lastname: Esmaeili
authors.affiliation: Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran, Iran; Chemical and Petroleum Engineering Department, Sharif University of Technology, Tehran, Iran. Electronic address: vosoughi@sharif.edu.
authors.forename: Manouchehr
authors.initials: M
authors.lastname: Vossoughi
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry; D055910:Antibodies, Immobilized / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D055664:Electrochemical Techniques / Q000379:methods*; D051379:Mice; D057139:Nanofibers / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D055550:Protein Stability; D013450:Sulfones / Q000737:chemistry*
chemical_list: D000907:Antibodies, Bacterial; D055910:Antibodies, Immobilized; D011108:Polymers; D013450:Sulfones; C022840:polyether sulfone
keywords: Antibody; Covalent binding; Electrospinning; Immobilization; Nanofiber