Abstract

Improving Peptide-Multiplexing Technology for Lab-On-Chip Autoantibody Based Diagnostics

Andreas Weinhaeusel, Senior Scientist, Austrian Institute of Technology GmbH

When developing assays for screening and validation of TAA-based biomarkers, many technical details need to be considered for immobilization of the antigens, processing conditions, and assay qualification in general. These variables influence the resulting data and eventually impede comparison and correlation of data from different platforms. Transferring candidate tumor-markers derived from protein-microarray to peptide based analysis we are evaluating planar and bead-array with respect to analytical conditions and performance. Because lack of single-antigen-/antibody pairs qualifying the different analysis platforms and testing cross-platform reproducibility of highly multiplexed tumor-autoantibody profiles is additionally challenging. Therefore we have established experimental settings using series of crosswise-mixed human IgG samples for evaluation and comparison of bead-arrays and microarrays, different array surfaces and processing conditions. Correlation and other goodness of fit measures can then be used for the assessment of our highly multiplexed analysis-platforms with respect to analytical performance. We present our approach of assessing reproducibility, array quality and cross-platform reproducibility for microarrays and suspension arrays. Additionally, we highlight the importance of comparing relative changes instead of absolute changes for cross-platform comparison of data and i.e. for evaluation of cross-platform reproducibility.

(Auto-)antibody testing has a high potential for enabling minimal invasive and early diagnostics and is well suited for lab-on-chip testing. We provide multiplexing array technologies and a complete pipeline for enabling autoantibody biomarker development for integration of antigenic proteins and peptides in final assays.