Investigation of a failed DNA assay leads to identification of an unexpected contaminant in a commonly used chromatography buffer.

Autor: Kuhns ST; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Semin DJ; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Soice NP; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Kappes A; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Wong HE; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Hwang TL; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Soto RJ; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Csordas AT; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Wu A; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Martinez-Cano D; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Babayan M; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Pelayo Y; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Hammond MR; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Nashed-Samuel Y; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA., Goudar CT; Attribute Sciences, Process Development, Drug Substance Technologies, Process Development, Quality Administration, Quality, Amgen Inc., Thousand Oaks, California, USA.
Jazyk: angličtina
Zdroj: Biotechnology progress [Biotechnol Prog] 2023 Mar; Vol. 39 (2), pp. e3307. Date of Electronic Publication: 2022 Nov 04.
DOI: 10.1002/btpr.3307
Abstrakt: For mammalian cell-derived recombinant biotherapeutics, controlling host cell DNA levels below a threshold is a regulatory requirement to ensure patient safety. DNA removal during drug substance manufacture is accomplished by a series of chromatography-based purification steps and a qPCR-based analytical method is most used to measure DNA content in the purified drug substance to enable material disposition. While the qPCR approach is mature and its application to DNA measurement is widespread in the industry, it is susceptible to trace levels of process-related contaminants that are carried forward. In this study, we observed failures in spike recovery studies that are an integral component of the qPCR-based DNA testing, suggesting the presence of an inhibitory compound in the sample matrix. We generated hypotheses around the origin of the inhibitory compound and generated multiple sample matrices and deployed a suite of analytical techniques including Raman and NMR spectroscopy to determine the origin and identity of the inhibitory compound. The caustic wash step and depth filter extractables were ruled out as root causes after extensive experimentation and DNA testing. Subsequently, 2-(N-morpholino)ethanesulfonic acid (MES), a buffer used in the chromatography unit operations, was identified as the source of the contaminant. A 500-fold concentration followed by Raman and NMR spectroscopy analysis revealed the identity of the inhibitory compound as polyvinyl sulfone (PVS), an impurity that originates in the MES manufacturing process. We have implemented PVS concentration controls for incoming MES raw material, and our work highlights the need for rigor in raw material qualification and control.
(© 2022 American Institute of Chemical Engineers.)
Databáze: MEDLINE
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