Current sensor platforms for detection/diagnosis of biothreat agent exposure depend on the use of antibodies to recognize and bind to specific antigens. To date, the selection of antibodies for inclusion in a final assay format has primarily relied on an antibody's performance in an enzyme-linked immunosorbant assay (ELISA) with little regard for quantifying the full spectrum of variables affecting antibody-antigen interaction. We recently instituted a quality program for the standardization of test methods for more fully characterizing and comparing the physical and functional properties of antibody reagents. The development and standardization of antibody testing provides an invaluable platform for the provision of consistent, high quality assays and reagents for both current biodetection platforms and development/validation of future systems. To this end, we report here the result of multiparametric studies of a variety of antibodies including: an scFV, Fab, mouse IgG, and human IgG. Each antibody was prescreened for both concentration and purity prior to analysis by ELISA, surface plasmon resonance (SPR), differential scanning calorimetry (DSC), dynamic light scattering (DLS) and biolayer interferometry (BLI). Antibodies were challenged by thermal denaturation or by exposure to a panel of interferents and binding patterns were observed. The results of this study provide not only standardized parametric data on biophysical properties, stability, and performance of each antibody, but also contribute to the development of a decisional analysis tool for expanding the confidence level of selecting antibody-based reagents that optimize field operational and performance metrics in future detection/diagnostic detection platforms.

Standardized platform for multiparametric characterization of antibody performance and stability

247th National Meeting (2014)

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