title: Surface Plasmon Resonance Biosensor for Detection of Bacillus anthracis, the Causative Agent of Anthrax from Soil Samples Targeting Protective Antigen.
abstract: Bacillus anthracis, the causative agent of anthrax is one of the most important biological warfare agents. In this study, surface plasmon resonance (SPR) technology was used for indirect detection of B. anthracis by detecting protective antigen (PA), a common toxin produced by all live B. anthracis bacteria. For development of biosensor, a monoclonal antibody raised against B. anthracis PA was immobilized on carboxymethyldextran modified gold chip and its interaction with PA was characterized in situ by SPR and electrochemical impedance spectroscopy. By using kinetic evaluation software, KD (equilibrium constant) and Bmax (maximum binding capacity of analyte) were found to be 20 fM and 18.74, respectively. The change in Gibb's free energy (∆G = -78.04 kJ/mol) confirmed the spontaneous interaction between antigen and antibody. The assay could detect 12 fM purified PA. When anthrax spores spiked soil samples were enriched, PA produced in the sample containing even a single spore of B. anthracis could be detected by SPR. PA being produced only by the vegetative cells of B. anthracis, confirms indirectly the presence of B. anthracis in the samples. The proposed method can be a very useful tool for screening and confirmation of anthrax suspected environmental samples during a bio-warfare like situation.
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: N
authors.initials: N
authors.lastname: Ghosh
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: G
authors.initials: G
authors.lastname: Gupta
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: M
authors.initials: M
authors.lastname: Boopathi
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: V
authors.initials: V
authors.lastname: Pal
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: A K
authors.initials: AK
authors.lastname: Singh
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: N
authors.initials: N
authors.lastname: Gopalan
authors.affiliation: Biotechnology Division, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002 India.
authors.forename: A K
authors.initials: AK
authors.lastname: Goel
keywords: Bacillus anthracis; Biosensor; Monoclonal antibody; Protective antigen; Surface plasmon resonance