Holistic analytical characterization and risk assessment of residual host cell protein impurities in an active pharmaceutical ingredient synthesized by biocatalysts. Issue 8 (4th May 2022)

• Record Type: Journal Article
• Authors: Wang, Fengqiang; Li, Xuanwen; Swanson, Michael; Guetschow, Erik; Winston, Matthew; Smith, Joseph P.; Hoyt, Erik; Liu, Zhijian; Richardson, Douglas; Bu, Xiaodong; Jawa, Vibha; Variankaval, Narayan
• Is Part Of: Biotechnology and bioengineering. Volume 119:Issue 8(2022)
• Page Start: 2088
• Page End: 2104
• Publication Date: 2022-05-04
• Subjects: analytical control strategy -- biocatalysis -- cGAS -- immunogenicity -- MK‐1454 -- residual host cell proteins -- risk assessment; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.28112
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Host cell proteins (HCPs) are a significant class of process‐related impurities commonly associated with the manufacturing of biopharmaceuticals. However, due to the increased use of crude enzymes as biocatalysts for modern organic synthesis, HCPs can also be introduced as a new class of impurities in chemical drugs. In both cases, residual HCPs need to be adequately controlled to ensure product purity, quality, and patient safety. Although a lot of attentions have been focused on defining a universally acceptable limit for such impurities, the risks associated with residual HCPs on product quality, safety, and efficacy often need to be determined on a case‐by‐case basis taking into consideration the residual HCP profile in the product, the dose, dosage form, administration route, and so forth. Here we describe the unique challenges for residual HCP control presented by the biocatalytic synthesis of an investigational stimulator of interferon genes protein agonist, MK‐1454, which is a cyclic dinucleotide synthesized using Escherichia coli cell lysate overexpressing cyclic GMP–AMP synthase as a biocatalyst. In this study, a holistic characterization of residual protein impurities using a variety of analytical tools including nanoscale liquid chromatography coupled to tandem mass spectrometry, together with in silico immunogenicity prediction of identified proteins, facilitated risk assessment and guided process development to achieve adequate removal of residual protein impurities in MK‐1454 active pharmaceutical ingredient. Abstract : Holistic characterization of residual protein impurities using a variety of analytical tools including enzyme‐linked immunosorbent assay, one‐dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis/Western blot, and in‐gel digestion coupled with nanoscale liquid chromatography coupled to tandem mass spectrometry was performed in a biocatalytically synthesized Merck investigational stimulator of interferon genes agonist, MK‐1454. This holistic analytical characterization together with in silico immunogenicity prediction of identified proteins facilitated risk assessment and guided process development to achieve adequate removal of residual protein impurities in the improved workup workflow for MK‐1454 biocatalysis and active pharmaceutical ingredient isolation.