title: Surface plasmon resonance immunosensor for the detection of Salmonella typhi antibodies in buffer and patient serum.
abstract: Surface plasmon resonance (SPR) immunosensor using 4-mercaptobenzoic acid (4-MBA) modified gold SPR chip was developed first time for the detection of flagellin specific antibodies of Salmonella typhi (S. typhi). Flagellin protein of S. typhi was prepared by recombinant DNA technology. The modification of gold chip with 4-MBA was in-situ characterized by SPR and electrochemical impedance spectroscopy. By using kinetic evaluation software, K(D) and B(max) values were calculated and found to be 26.3 fM and 62.04 m°, respectively, for the immobilized monoclonal antibody (Moab) of recombinant flagellin (r-fla) protein of S. typhi (r-fla S. typhi). In addition, thermodynamic parameters such as ΔG, ΔH and ΔS were determined first time for r-fla S. typhi and Moab of r-fla S. typhi interactions and the values revealed the interaction between r-fla S. typhi and Moab of r-fla S. typhi as spontaneous, endothermic and entropy driven one. Moreover, healthy human serum samples and patient sera (Widal positive and Widal negative) were subjected to SPR analysis. The present SPR based approach provides an alternative way for S. typhi detection in less than 10 min.
authors.affiliation: Defence Research and Development Establishment, DRDO, Gwalior 474002, India.
authors.forename: Garima
authors.initials: G
authors.lastname: Gupta
authors.forename: P K
authors.initials: PK
authors.lastname: Sharma
authors.forename: B
authors.initials: B
authors.lastname: Sikarwar
authors.forename: S
authors.initials: S
authors.lastname: Merwyn
authors.forename: S
authors.initials: S
authors.lastname: Kaushik
authors.forename: M
authors.initials: M
authors.lastname: Boopathi
authors.forename: G S
authors.initials: GS
authors.lastname: Agarwal
authors.forename: Beer
authors.initials: B
authors.lastname: Singh
mesh_terms: D000911:Antibodies, Monoclonal / Q000097:blood / Q000737:chemistry; D001565:Benzoates / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D058266:Dielectric Spectroscopy / Q000379:methods; D005408:Flagellin / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D012485:Salmonella typhi / Q000276:immunology / Q000302:isolation & purification*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D001565:Benzoates; D011994:Recombinant Proteins; D013438:Sulfhydryl Compounds; C013594:4-mercaptobenzoate; D005408:Flagellin