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