Process-related impurity analysis of biotherapeutics

Gyrolab^® systems and immunoassays speed up the bioprocessing workflow

Process-related impurity concentrations are highly regulated in the manufacture of biologic drugs because of their immunogenic potential. Bioanalytical methods to detect and measure process related impurities must be developed carefully to characterize heterogeneous mixtures of host cell proteins (HCPs) present in samples, or the copurification of culture or process impurities such as protein A together with the biotherapeutic.

Heterogeneous mixtures of host cell proteins (HCPs), for example
Chinese Hamster Ovary (CHO)-HCP, E. coli HCP, phospholipase B-like 2 (PLBL-2) and transferrin
Process-specific impurities
Carryover of culture or process impurities such as Protein A and insulin

Impurity analysis using current ELISA technologies has limitations that include labor-intensive protocols, lower throughput and narrow dynamic range. As a result, repeat analysis is often required, which further delays project decisions and lot approvals. Automating impurity analysis with the miniaturized, microfluidic immunoassays supported by Gyrolab systems shortens turn-around time, broadens dynamic ranges, and delivers reproducible, high-quality data, with fewer manual operations. These benefits make Gyrolab systems an ideal technology to support all phases of bioprocess and production of biotherapeutics.

Gyrolab immunoassays provide faster time to data compared to traditional ELISA methods (data courtesy of Medimmune, the global biologics research and development arm of AstraZeneca).

HCP analysis with expanded dynamic range

One primary goal during HCP assay development is a broad dynamic range to avoid repeat analysis and additional sample dilutions. A broad dynamic range is especially important for bioprocess HCP analysis, since the concentration range of HCP impurities in process samples may vary greatly. Gyrolab assays and kits offer broad analytical ranges, reducing the the number of dilutions required to hit the analytical range, thereby reducing the need for repeat runs. CHO HCP 3G Solution for Gyrolab^® consists of the CHO HCP 3G Assay Reagent Set for Gyrolab from Cygnus Technologies and the Gyrolab Bioaffy ™ 1000 HC Assay Toolbox. The reagent set is derived from the same antibodies and antigen used in the Cygnus 3rd Generation CHO HCP ELISA kit, Cat# F550-1.

Gyrolab CHO-HCP offers shows a broad analytical range of approximately 2 – 8000 ng/mL compared to 1 – 1000 ng/mL for ELISA.

In situations where assays with process-specific reagents are required, Gyrolab Bioaffy™ 1000 HC Assay Toolbox and Gyrolab Bioaffy™ 4000 HC Assay Toolbox provide the flexibility and automation needed to deliver rapid results.

Protein A and culture impurities

Other bioprocess impurities may carry over from purification columns or culture media. Gyrolab Protein A Kit detects GE Healthcare MabSelect SuRe™ ligands in samples, and carefully tested Gyrolab assay protocols are available for the analysis of Human Transferrin, and Insulin impurities. The broad analytical range of Gyrolab immunoassays provides several logs of additional working range expansion over ELISA and a lower minimum required dilution (MRD).