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Quantitative analysis of factor P (Properdin) in monkey serum using immunoaffinity capturing in combination with LC-MS/MS

Gao, Xinliu and Lin, Hui and Krantz, Carsten and Garnier, Arlette and Flarakos, Jimmy and Tse, Francis and Li, Wenkui (2016) Quantitative analysis of factor P (Properdin) in monkey serum using immunoaffinity capturing in combination with LC-MS/MS. Bioanalysis.

Abstract

Factor P (Properdin) is an endogenous protein found in the serum of human and other animals. As the only known positive regulator of the complement system, factor P plays a key role in innate immune defense against many diseases. Quantification of factor P in serum samples is important in understanding the pharmacodynamics (PD) of drug candidates. However, developing a suitable quantification method is challenging because of the endogenous nature of the protein. In the present work, a sensitive and selective method combining immunoaffinity capturing and LC-MS/MS has been developed and validated for the first time for quantitative analysis of factor P in monkey serum. True monkey serum was used for the preparation of calibration standards and QCs. The method was linear and rugged over a dynamic range of 125 to 25000 ng/mL using 50 uL sample volume. The intra- and inter-run precision (%CV) of the QC sample results were within 7.2% (CV) and the accuracy (%Bias) within ±16.8% across all QC concentrations evaluated. Other validation parameters, including stability under various conditions, extraction recovery and matrix effect, all met the acceptance criteria. The validated method was successfully implemented for the quantitative analysis of factor P in monkey serum samples in support of a preclinical PD study to assess response of a drug candidate.

Factor P (Properdin), Immunoaffnity capturing, LC-MS/MS, monkey serum

Item Type: Article
Date Deposited: 27 Apr 2016 23:45
Last Modified: 27 Apr 2016 23:45
URI: https://oak.novartis.com/id/eprint/26508

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