Answer: We recommend working with generic ELISA kits during early drug development phases. The proteins of host cells are specific for a given process and can vary, depending on factors like temperature and media. Most generic HCP assay providers offer one kit per cell line that should be suitable for all processes. At BioGenes, you have the possibility to identify the most suitable generic HCP ELISA kit for your process by our enhanced 4-kit HCP ELISA approach for E. coli and CHO, each using a different set of anti-HCP antibodies. Our scientists at BioGenes can assist you in identifying the most suitable generic HCP ELISA kit for your process. This performance study evaluates our four kit types (optionally, we can include generic HCP ELISA kits from other manufacturers) and is separated into two parts. Ideally, the customer provides a panel of in-process controls (IPCs) from different purification steps.
In the first part of the performance study, we will examine the provided samples by:
| Coverage Testing | The coverage of the four kits will be tested by 1D Western Blot and immunostaining with antibodies from the respective 360-HCP ELISA kit to choose a preferred kit. |
| Determine HCP content | Samples from different purification steps (e.g. three IPCs) are analyzed by ELISA to monitor the decrease of HCPs. If no clear reduction is observed, the generic ELISA kit is unsuitable for the specific manufacturing process. No reduction of HCP level can be evidence for cross-reactivity of the antibodies with the recombinant drug substance due to false-positive assay signals. |
| ELISA accuracy | To verify the ELISA accuracy, we test the most suitable selected 360-HCP ELISA kit with our spike-and-recovery protocol. If available, a process-related mock sample can be used to confirm the results. |
In the second part, a comprehensive analysis of a single process sample will be done based on the results from the first part of the performance study. The HCP coverage of the selected generic HCP ELISA kit will be determined by classical 2D Western Blot and confirm the suitability of the chosen kit for your process. Optionally, BioGenes offers immunoaffinity chromatography (IAC) followed by 2D DIGE analysis as an orthogonal method to determine the coverage of the selected kit under almost native assay conditions.
The results of each part of the performance study will be communicated to our customers in a detailed report. Get in touch with us to learn more about the performance study or our products!