ELISA on a chip: developing a novel antigen microarray assay for high-throughput ELISA assays

Shlomia Levy ^1,2 Lilach M. Friedman ^1,2 Lester M. Shulman ³ Tomer Hertz Tomer Hertz ^1,2,4

¹The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Israel
²The National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, Israel
³Dept. of Epidemiology and Preventive Medicine,school of Public Health, Sackler Faculty of Medicine, Tel Aviv University, Israel
⁴Vaccine and Infectious Disease Division, Fred Hutch Cancer Research Center, USA

ELISA is a plate-based assay for detecting and quantifying antibodies etc. The assay is widely in many other research fields. Antigen microarrays (AMs) are a high throughput Ab binding assay that allows the quantification of Ab responses to thousands of antigens, including proteins, peptides, and whole viruses.

Here we developed a novel AM based ELISA assay. Our assay has several clear advantages over the ELISA assay: (1) It can be used to simultaneously detect antibody binding to thousands of different antigens; (2) It requires small volumes of serum samples; and (3) It can be easily used to profile over 100 subjects in a single experiment.

To test our novel assay we used a pilot array spotted with eight influenza Hemagglutinin (HA) proteins and one Nucleoprotein (NP) from different strains. Antigens were spotted in triplicates across serial dilutions. For comparisons we developed a low-volume 384 well plate traditional ELISA. We used serum samples from 10 adult subjects, as well as monoclonal antibodies to compare the two assays. Each sample was run on both assays at 15 serial dilutions ranging from 1:25-1: 1:409,600.

We found that the AM could be used to generate binding curves similar to the traditional ELISA, using both serial dilutions of the sample or the antigens. Overall correlations between the binding curves of the two assays were significant. Our results highlight the feasibility of using our novel ELISA on chip assay for rapid characterization of antibody binding as a low-volume high-throughput alternative to the traditional ELISA assay.