Real-time Monitoring of Antibody Quality Control using FabRICATOR

September 23, 2015 | Applications, References |

The production of antibody-based therapeutics requires careful monitoring of quality attributes. Researchers at Amgen developed a technology to model and monitor the high-mannose content of the antibody in real-time using FabRICATOR digestion (Zupke et al. 2015).

The amount of high-mannose glycans at the N-linked glycan site on therapeutic antibodies have been shown to impact antibody clearance and thus the pharmacokinetics of the antibody. To follow and control the amount of high-mannose glycan in the production of an IgG1 antibody, researchers at Amgen developed a technology based on FabRICATOR for near real-time LC-MS analysis.

In the paper, Zupke et al. developed mathematical models to predict the amount of high-mannose residues on the antibody as a function of added mannose sugar. To control the high-mannose glycan in real-time, FabRICATOR was used directly on filtered cell-culture media. The antibody fragments were reduced and the 25 kDa fragments analyzed using LC-MS.

The traditional approach to study IgG glycosylation involves release of glycans using PNGaseF, labeling with a fluorescent dye and separation of the glycan structures using HILIC chromatography. Due to the time for sample preparation in this approach, it is not well suited for on-line monitoring. Instead, FabRICATOR was incubated with cell culture medium for 30 min, followed by reduction for 10 min, and then separation and analysis using LC-MS. This allowed a close to real-time response of the quality of the batch and monitoring of the high-mannose content.

Zupke and colleagues at Amgen presents an impressive study that highlights the importance of controlling critical quality attributes during the process development, that could avoid extensive product characterization.