Reduction of incubation times for viral pathogen immunoassays by using elevated temperatures as monitored by atomic force microscopy

ANYL 173

Karen M. Kwarta, kstrangh@iastate.edu1, Jeremy D. Driskell, driskell@iastate.edu1, Julia F. Ridpath, jridpath@nadc.ars.usda.gov2, John D. Neill, jneill@nadc.ars.usda.gov2, and Marc D. Porter, mporter@porter1.ameslab.gov1. (1) Department of Chemistry, Iowa State University, 44 Spedding Hall, Ames, IA 50010, (2) National Animal Disease Center, United States Department of Agriculture, Ames, IA 50010
This presentation describes the use of elevated temperatures to reduce the time needed to selectively extract viruses from a sample solution onto a solid capture substrate coated with monoclonal antibodies (Mab) specific for the viruses. Raising incubation temperatures causes a change in the antibody-virus interaction and in the rate of mass transport for the virus to the substrate. Porcine parvovirus (PPV) and feline calicivirus (FCV) were chosen as model targets because their well defined size and shape enables facile enumeration by imaging with atomic force microscopy (AFM). This readout approach yielded a label-free assay, which was used to study binding kinetics. The binding kinetics of both viruses to their respective Mab's was examined at two different temperatures (25 °C and 37 °C). Dose-response curves show that the extraction time can be reduced to 1 h at 37 °C while only reducing the detection limit achieved with 16 h incubation at 25 °C by a factor of two.
 

General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006