title: Development of automated amperometric detection of antibodies against Bacillus anthracis protective antigen.
abstract: Picogram levels of antibodies against the protective antigen (PA) of Bacillus anthracis were detected in an automated electrochemical sandwich-type enzyme-linked immunosorbant assay. The antibodies were captured and detected using an 8 x 3 array of 50-microm-diameter cavities. The reagent and sample volumes were as low as 200 nL in a less than 25-min assay from capture to signal generation. The electrochemical detection of the antibodies was demonstrated at 0.05-10 microg/mL containing only 10-5,000 pg antibodies. The limit of detection is 10 fg for a 200-nL sample. Detection of anti-PA immunoglobulin G performed in spiked normal human serum and fresh whole human blood did not show a significant difference from detection in a buffer. The initial automation of the assay involved the use of a digital syringe pump for the delivery of reagents to the capture surface.
authors.affiliation: Vegrandis, LLC, 700 W. Research Ctr. Blvd., ENRC 1208, Fayetteville, AR 72701, USA. zoraida.aguilar@vegrandis.com
authors.forename: Zoraida P
authors.initials: ZP
authors.lastname: Aguilar
authors.forename: Mali
authors.initials: M
authors.lastname: Sirisena
mesh_terms: D000907:Antibodies, Bacterial / Q000032:analysis*; D000942:Antigens, Bacterial / Q000276:immunology*; D001331:Automation; D001427:Bacterial Toxins / Q000276:immunology*; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D006801:Humans; D012680:Sensitivity and Specificity
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; C030325:anthrax toxin