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Activity Number: 463 - Topics in Biostatistics
Type: Contributed
Date/Time: Thursday, August 6, 2020 : 10:00 AM to 2:00 PM
Sponsor: Section on Physical and Engineering Sciences
Abstract #313928
Title: Applications of Design of Experiments to Develop a Robust Assay for Replacing the Rabbit Blood Sugar Bioidentity Test of Insulin Glargine
Author(s): Monisha Dey* and Junming Yie
Companies: Merck & Co., Inc and Merck & Co., Inc
Keywords: functional cell-based assay; robustness; USP<121>; comparability; qualification

A rabbit blood sugar bioidentity assay is required by the FDA to evaluate biological activity for all insulin and its analogs per USP< 121> guideline. Not only are a large number of live animals used, but the rabbit blood sugar method is also highly variable and expensive. Our goal is to develop a functional cell-based assay to replace rabbit blood sugar method. An H4IIE G6P-Luc reporter assay was developed by utilizing insulin’s role in regulating hepatic gluconeogenesis pathway. The cell-based assay was developed and optimized through design of experiments (DOE) to demonstrate good dose-dependent responsiveness to insulin. We further qualified the assay and demonstrated excellent performance characteristics of linearity, accuracy, and precision. A robustness study using split-plot DOE was then conducted to define critical factors for assay performance. We compared this newly developed assay with a previously established cell-based assay using orthogonal regression methodology and showed its advantages. In conclusion, we have developed a physiologically relevant and robust functional cell-based assay that is suitable to replace rabbit blood sugar method.

Authors who are presenting talks have a * after their name.

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