Application Note

Automate HEK 293 HCP Detection Using Antibody Reagents

Source: bio-techne
bio techne

Human Embryonic Kidney (HEK) 293 cells are often used as host cell lines for protein production or viral vector scale-up, as part of bio-therapeutic or cell and gene therapy manufacturing workflows. HEK 293 host cell proteins (HCPs) have a similar nature, risk profile, and manufacturing process optimization requirement as HCPs from other expression systems. As such, HEK HCPs need to be monitored and reduced at every step of the manufacturing workflow. Residual HCPs that go undetected can decrease product efficacy, alter its quality, and trigger immunotoxic reactions after administration of bio-therapeutics or cell and gene therapies3.

To meet increasing business demands and safety standards, fast and reproducible detection of HCPs is essential during viral vector and protein production. Accurate in-process reporting is complicated by the wide range of in-process matrices present throughout the purification process. The initial crude viral vector harvest may be obtained via the lysis or disruption of host cells, generating high levels of HCPs. In contrast, final purification stages contain significantly lower levels. You’ll need to carefully and extensively dilute samples at the early stages of purification to ensure detection within the dynamic range of your assay. The sensitivity of your assay should enable the detection of low levels of HCPs that may be present in the final purification stages. This workflow requires a quantitative solution that delivers both sensitivity and a broad dynamic range.

In this application note, we show how the Simple Plex™ HEK 293 HCP 3G Assay on the Ella™ platform consistently delivers sensitivity and a broad dynamic range. By pairing Simple Plex microfluidic immunoassay technology with the Cygnus Technologies HEK 293 HCP Resupply 1 polyclonal antibodies, we demonstrate utility for fast, accurate detection and quantitation of HEK HCP contaminants from viral vector and protein production samples.

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