HCP Coverage Analysis
- Evaluation and comparison of ELISAs
- Low amount of antibody required - 1 mg
- Alternative to 2D Gels & Western Blotting
Do you know how well your ELISA covers the HCPs in your sample?
When you need to evaluate the antibodies’ quality of a generic or process-specific HCP ELISA, we can help. We use advanced MS-based coverage analysis (ELISA-MS™) to identify each HCP the ELISA can measure. In collaboration with BioGenes, we also offer 2D DIGE and Immunoaffinity Chromatography ...
Do you know how well your ELISA covers the HCPs in your sample?
When you need to evaluate the antibodies’ quality of a generic or process-specific HCP ELISA, we can help. We use advanced MS-based coverage analysis (ELISA-MS™) to identify each HCP the ELISA can measure. In collaboration with BioGenes, we also offer 2D DIGE and Immunoaffinity Chromatography for coverage analysis.
For ELISA-MS, you can use an early process sample containing the drug substance, and you don’t need to develop a null cell lysate from a mock fermentation. The host cell proteins in the sample are immunocaptured using the ELISA antibodies and then identified by LC-MS/MS. As a result, you get the list of the HCPs recognized by the ELISA antibodies.
How does ELISA-MS™ analysis compare to 2D PAGE and Western blot?
ELISA-MS™ is a proven method recognized as a better alternative to spot counting in 2D gel electrophoresis and Western blot because it provides more detailed and accurate information about each HCP. An Immunocapture LC-MS analysis (ELISA-MS™) will thus expand your knowledge of the HCP-ELISA’s specificity and applicability to your biopharmaceutical. It may also reduce the risk of project delays due to poorly documented HCP ELISA.
A standard ELISA-MS™ HCP coverage analysis includes:
- ELISA setup for optimal antibody-HCP binding.
- Immunocapture with ELISA polyclonal antibodies.
- Identification of both total HCPs and affinity-purified HCPs by sensitive LC-MS/MS.
- Reporting of HCPs covered by antibodies and also a list of total HCPs in the mock lysate or early process sample.
2D DIGE and Immunoaffinity Chromatography
Current guidelines recommend coverage determination using several orthogonal methods. In collaboration with BioGenes, we offer different types of ELISA coverage methods. These include Immunoaffinity Chromatography IAC, combined with either A) 2D DIGE comparative analysis of spot patterns for HCP Coverage percentage determination, or B) LC-MS for identification of IAC captured HCPs, as orthogonal methods for HCP coverage analysis.
Send us a description of your project and we will be glad to discuss it with you.
