Detection of Residual Enzymatic Activity in Plant-Derived Recombinant Human Serum Albumin Products Using 31P NMR Spectroscopy

Published

December 2, 2014

Author(s)

Robert G Brinson, John P. Marino, Zvi Kelman, Gary Giulian

Abstract

The minimization of impurities in biotherapeutics, such as host cell proteins (HCPs) and other molecules, is critical due to potential toxicity and/or immunogenic responses to foreign-sourced molecules in the human body. Currently, the ELISA assay, due to its high sensitivity and dynamic range, is the standard for detection of residual HCP contamination in recombinantly-produced biologics from bacterial, yeast, or mammalian expression systems1. Alternatively, two-dimensional liquid chromatography coupled to mass spectrometry (2D-LCMS) is being developed as a tool for assessing this critical quality attribute2, 3. Both methods rely on the direct detection of HCPs. In the case of HCP enzymes, the mass level of contamination may fall below the threshold of detection using these methods and underestimate the true nature of the contamination. To address this point, here we demonstrate a sensitive, label-free nuclear magnetic resonance spectroscopy

Citation

Analytical Chemistry

Volume

86

Issue

23

Pub Type

Journals

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DOI Link

Keywords

host cell protein, phytase, phytic acid, ATP, 31P NMR

Citation

, R. , Marino, J. , Kelman, Z. and Giulian, G. (2014), Detection of Residual Enzymatic Activity in Plant-Derived Recombinant Human Serum Albumin Products Using 31P NMR Spectroscopy, Analytical Chemistry, [online], https://doi.org/10.1021/ac503864m (Accessed April 23, 2024)