title: Antibody immobilization using pneumatic spray: comparison with the avidin-biotin bridge immobilization method.
abstract: The formation of a thin antibody film on a glass surface using pneumatic spray was investigated as a potential immobilization technique for capturing pathogenic targets. Goat-Escherichia coli O157:H7 IgG films were made by pneumatic spray and compared against the avidin-biotin bridge immobilized films by assaying with green fluorescent protein (GFP) transformed E. coli O157:H7 cells and fluorescent reporter antibodies. Functionality, stability, and immobilization of the films were tested. The pneumatic spray films had lower fluorescence intensity values than the avidin-biotin bridge films but resulted in similar detection for E. coli O157:H7 at 10(5)-10(7)cells/ml sample concentrations with no detection of non-E. coli O157:H7 strains. Both methods also resulted in similar percent capture efficiencies. The results demonstrated that immobilization of antibody via pneumatic spray did not render the antibody non-functional and produced stable antibody films. The amount of time necessary for immobilization of the antibody was reduced significantly from 24h for the avidin-biotin bridge to 7 min using the pneumatic spray technique, with additional benefits of greatly reduced use of materials and chemicals. The pneumatic spray technique promises to be an alternative for the immobilization of antibodies on glass slides for capturing pathogenic targets and use in biosensor type devices.
authors.affiliation: Department of Chemistry, University of South Florida, 4202 E. Fowler Ave., Tampa, FL 33620-5250, USA.
authors.forename: Jhon
authors.initials: J
authors.lastname: Figueroa
authors.forename: Sonia
authors.initials: S
authors.lastname: Magaña
authors.forename: Daniel V
authors.initials: DV
authors.lastname: Lim
authors.forename: Rudy
authors.initials: R
authors.lastname: Schlaf
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry*; D055910:Antibodies, Immobilized / Q000737:chemistry*; D001360:Avidin / Q000737:chemistry; D015374:Biosensing Techniques; D023482:Bioterrorism; D001710:Biotin / Q000737:chemistry; D003362:Cost-Benefit Analysis; D019453:Escherichia coli O157 / Q000235:genetics / Q000276:immunology; D005453:Fluorescence; D005516:Food Microbiology; D005898:Glass / Q000737:chemistry*; D006041:Goats; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D013499:Surface Properties; D013997:Time Factors
chemical_list: D000907:Antibodies, Bacterial; D055910:Antibodies, Immobilized; D007074:Immunoglobulin G; D001360:Avidin; D049452:Green Fluorescent Proteins; D001710:Biotin