Polymethacrylate coated electrospun PHB fibers: An exquisite outlook for fabrication of paper-based biosensors.

Electrospun polyhydroxybutyrate (PHB) fibers were dip-coated by polymethyl methacrylate-co-methacrylic acid, poly(MMA-co-MAA), which was synthesized in different molar ratios of the monomers via free-radical polymerization. Fabricated platfrom was employed for immobilization of the dengue antibody and subsequent detection of dengue enveloped virus in enzyme-linked immunosorbent assay (ELISA). There is a major advantage for combination of electrospun fibers and copolymers. Fiber structre of electrospun PHB provides large specific surface area available for biomolecular interaction. In addition, polymer coated parts of the platform inherited the premanent presence of surface carboxyl (-COOH) groups from MAA segments of the copolymer which can be effectively used for covalent and physical protein immobilization. By tuning the concentration of MAA monomers in polymerization reaction the concentration of surface -COOH groups can be carefully controlled. Therefore two different techniques have been used for immobilization of the dengue antibody aimed for dengue detection: physical attachment of dengue antibodies to the surface and covalent immobilization of antibodies through carbodiimide chemistry. In that perspective, several different characterization techniques were employed to investigate the new polymeric fiber platform such as scanning electron microscopy (SEM), atomic force microscopy (AFM), water contact angle (WCA) measurement and UV-vis titration. Regardless of the immobilization techniques, substantially higher signal intensity was recorded from developed platform in comparison to the conventional ELISA assay.

[1]  E. Alocilja,et al.  Surface functionalization of electrospun nanofibers for detecting E. coli O157:H7 and BVDV cells in a direct-charge transfer biosensor. , 2010, Biosensors & bioelectronics.

[2]  Fatimah Ibrahim,et al.  Recent advances in surface functionalization techniques on polymethacrylate materials for optical biosensor applications. , 2014, The Analyst.

[3]  F. Ibrahim,et al.  Synthesis and characterization of methacrylic microspheres for biomolecular recognition: Ultrasensitive biosensor for Dengue virus detection , 2014 .

[4]  Wei Li,et al.  Preparation and characterization of bovine serum albumin surface-imprinted thermosensitive magnetic polymer microsphere and its application for protein recognition. , 2014, Biosensors & bioelectronics.

[5]  F. Ibrahim,et al.  Synthesis and processing of ELISA polymer substitute: The influence of surface chemistry and morphology on detection sensitivity , 2014 .

[6]  P. Supaphol,et al.  Bone scaffolds from electrospun fiber mats of poly(3-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and their blend , 2007 .

[7]  B. R. Coad,et al.  Controlled covalent surface immobilisation of proteins and peptides using plasma methods , 2013 .

[8]  Wenbo Lu,et al.  A novel label-free amperometric immunosensor for carcinoembryonic antigen based on Ag nanoparticle decorated infinite coordination polymer fibres. , 2014, Biosensors & bioelectronics.

[9]  F. Ibrahim,et al.  Polymethyl methacrylate-co-methacrylic acid coatings with controllable concentration of surface carboxyl groups: A novel approach in fabrication of polymeric platforms for potential bio-diagnostic devices , 2014 .

[10]  Dietmar W. Hutmacher,et al.  Design, fabrication and characterization of PCL electrospun scaffolds—a review , 2011 .

[11]  Alankar Shrivastava,et al.  Methods for the determination of limit of detection and limit of quantitation of the analytical methods , 2011 .

[12]  V. Deubel,et al.  Enzyme-Linked Immunosorbent Assay Specific to Dengue Virus Type 1 Nonstructural Protein NS1 Reveals Circulation of the Antigen in the Blood during the Acute Phase of Disease in Patients Experiencing Primary or Secondary Infections , 2002, Journal of Clinical Microbiology.

[13]  P. Allongue,et al.  Semiquantitative study of the EDC/NHS activation of acid terminal groups at modified porous silicon surfaces. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[14]  E. Hunsperger,et al.  Comparison of Two Commercially Available Dengue Virus (DENV) NS1 Capture Enzyme-Linked Immunosorbent Assays Using a Single Clinical Sample for Diagnosis of Acute DENV Infection , 2008, Clinical and Vaccine Immunology.

[15]  Jyh-Ping Chen,et al.  Fabrication of electrospun poly(methyl methacrylate) nanofibrous membranes by statistical approach for application in enzyme immobilization , 2009 .

[16]  Chien-Chong Hong,et al.  Point-of-care protein sensing platform based on immuno-like membrane with molecularly-aligned nanocavities. , 2013, Biosensors & bioelectronics.

[17]  K. Yuen,et al.  Serotype 1-Specific Monoclonal Antibody-Based Antigen Capture Immunoassay for Detection of Circulating Nonstructural Protein NS1: Implications for Early Diagnosis and Serotyping of Dengue Virus Infections , 2006, Journal of Clinical Microbiology.

[18]  Kazuhiko Ishihara,et al.  Electrospun phospholipid polymer substrate for enhanced performance in immunoassay system. , 2012, Biosensors & bioelectronics.

[19]  R. Césaire,et al.  Relationship between nonstructural protein 1 detection and plasma virus load in Dengue patients. , 2010, The American journal of tropical medicine and hygiene.

[20]  Ting-Hsiang Lin,et al.  Comparison of Capture Immunoglobulin M (IgM) and IgG Enzyme-Linked Immunosorbent Assay (ELISA) and Nonstructural Protein NS1 Serotype-Specific IgG ELISA for Differentiation of Primary and Secondary Dengue Virus Infections , 2003, Clinical Diagnostic Laboratory Immunology.

[21]  E. Szili,et al.  Polyoctanediol Citrate/Sebacate Bioelastomer Films: Surface Morphology, Chemistry and Functionality , 2010, Journal of biomaterials science. Polymer edition.

[22]  R. Kelly,et al.  Nanofibrous membranes for single-step immobilization of hyperthermophilic enzymes , 2014 .

[23]  C. Chu,et al.  Label-free detection of DNA using novel organic-based electrolyte-insulator-semiconductor. , 2010, Biosensors & bioelectronics.

[24]  S. Xiao,et al.  Different EDC/NHS activation mechanisms between PAA and PMAA brushes and the following amidation reactions. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[25]  C. Lim,et al.  Recent development of polymer nanofibers for biomedical and biotechnological applications , 2005, Journal of materials science. Materials in medicine.

[26]  M. Kotaki,et al.  Surface modified nonwoven polysulphone (PSU) fiber mesh by electrospinning : a novel affinity membrane , 2006 .

[27]  Jeong‐Yeol Yoon,et al.  Adsorption of BSA on Highly Carboxylated Microspheres—Quantitative Effects of Surface Functional Groups and Interaction Forces , 1996 .

[28]  L. Kubota,et al.  Immunospot assay based on fluorescent nanoparticles for Dengue fever detection. , 2013, Biosensors & bioelectronics.

[29]  Rong Huang,et al.  Plasmonic ELISA for the ultrasensitive detection of Treponema pallidum. , 2014, Biosensors & bioelectronics.

[30]  Y. Ikada,et al.  Introduction of functional groups onto the surface of polyethylene for protein immobilization. , 1993, Biomaterials.