Reliability and Reproducibility with Automated MIMIC® Assays

VaxDesign has extensively tested its automated assays for reliability and reproducibility. The results shown below demonstrate that the automated assays are superior to manual assays.

We often use a traditional (unfractionated) PBMC assay to ensure the quality of our blood products and as a benchmark in which to evaluate the utility of MIMIC® assays. Because of the simplicity of this assay, we felt it provided an excellent proof-of-concept demonstration of the capabilities of our automated systems to perform biological assays. To qualify the automated PBMC assay against a manual experiment, we performed this assay in parallel using the same cells and reagents.

The automated PBMC assay is comparable to the manual PBMC assay. Frozen PBMCs were thawed, counted, and dispensed in the presence or absence of a panel of stimulators, as indicated. After 24 hr, the culture supernatants were harvested and evaluated by protein microarray for the expression of a series of immunologically relevant cytokines. Each bar represents an average of three samples with a standard deviation value.

Fabrication of the PTE Module: No Change in Key Cytokines

Automated and manual PTE constructs elicit similar basal-level cytokine responses. Culture supernatants were collected 7 days after collagen cushions were seeded with HUVEC cells and then evaluated by bioplex analysis. Each sample represents an average response from 5 separate assays containing 4 pooled samples. The error bars show standard deviation values.

The next figure shows a consolidated graph directly comparing influenza-specific IgG concentrations obtained by the manual and automated ELISA assays. Importantly, while the automated and manual methods generated nearly identical average values, the automated method showed a lower degree of error and required nearly 80% less labor hours than the manual approach. As evident from this and other assays, the well-to-well and plate-to-plate variation of the automated method is extremely low, indicating the robotic platform can provide a high degree of inter- and intra-assay consistency. We believe this is an important aspect for reproducibility and reliability of not only the MIMIC® System, but also other immunological assays.