Improving ELISA Assay Efficiency in Pharmacokinetic Studies

• Activity Number: 437
• Type: Contributed
• Date/Time: Tuesday, August 11, 2015 : 2:00 PM to 3:50 PM
• Sponsor: Biopharmaceutical Section
• Author(s): Anna Decker* and William Forrest and Hanine Anezinos and Kelly Loyet and Moulay Hicham Alaoui Ismaili
• Companies: and Genentech, Inc. and Genentech, Inc. and Genentech, Inc. and Genentech, Inc.
• Keywords: pharmacokinetic studies ; ELISA

Abstract

The enzyme-linked immunosorbent assay (ELISA) is a widely used analytical method to quantify biotherapeutics in pharmacokinetic (PK) studies. It allows for accurate concentration determination of a therapeutic protein in various animal matrices. Sample concentrations may vary widely, which necessitates serial dilutions to quantify unknown concentrations within a limited standard curve range. A larger number of serial dilutions uses more consumables and is more labor-intensive and time consuming for the analysts. Therefore, there is substantial interest in improving the efficiency of these assays. Using historical data from pharmacokinetic studies, we investigated the impact of reducing the number of dilutions in each dilution series and assessed the utility of an automatic flagging procedure for non-linear dilutions. We found that a 25% reduction in the number of dilutions maintained sample quality and that an automatic flagging procedure could be a useful alternative to manual exclusion of non-linear dilutions via the analyst.