Services ELISA characterization and coverage analysis

ELISA reagent characterization and HCP coverage analysis

This page has been reviewed and verified by Ejvind Mortz, PhD in Protein Chemistry & Molecular Biology

When you need a detailed characterization of your HCP-ELISA antibodies, advanced ELISA-MS™ is the best technology available. Unlike other methods, ELISA-MS™ analysis utilizes immunocapture directly in the ELISA well and measures the actual protein-antibody binding using LC-MS without artificial systems, thus mimicking the actual conditions of the ELISA for evaluating the HCP coverage.

With ELISA-MS™, you can accurately evaluate the quality of antibodies in any commercial, platform, and process-specific kit and identify the individual Host Cell Proteins (HCPs) measured by the ELISA antibodies - and also the HCPs that are not measured.

This detailed characterization of ELISA antibodies enables you to evaluate and select the best-fit ELISA for monitoring the product- or process-related impurities in your product. It also reduces the risk that abundant or potentially problematic HCPs go undetected by your ELISA.

Many of our clients have successfully used ELISA-MS™ data to document the suitability of commercial and process-specific assays, including it as part of IND and BLA submissions.

Why use LC-MS for ELISA characterization?

Compare ELISAs and document HCP coverage

Document coverage percentage, identify covered vs. uncovered HCPs, and get a list of high-risk HCPs – enabling you to select the best fit ELISA for your product

Bridging studies

Characterize ELISA reagents and HCP standards to assess why the ELISA shows dilutional nonlinearity or inconsistent results

Troubleshooting ELISAs

Investigate issues caused by antibody/ drug substance cross-reactivity, insufficient specificity, incorrect standards for the calibration curve, etc.

Is your ELISA giving different results - or not showing dilutional linearity?

Many of our clients face the challenge that different ELISA kits, even from the same vendor, can show very different results — sometimes more than 10 to 100-fold in- or decreases in HCP amounts — when analyzing identical batches.

Using LC-MS, we can uncover if the inconsistent results may be due to an unintended change in the HCP standard. Such a change can make it a poor fit for your drug sample or antibodies, resulting in a different HCP concentration readout even with the same HCP antibodies. Other reasons for the inconsistent results, which ELISA-MS™ can uncover, might be that a new generation of antibodies binds better — or worse — to the proteins in your sample.

Have you convincingly documented that ELISA covers most Host Cell Proteins in your drug substance?

Regulatory authorities are concerned that abundant HCPs in a drug product may go undetected by ELISA or that your drug substance may contain undetected HCPs of concern that impact patient safety or drug stability.

ELISA-MS™ data provides not only the coverage percentage of your ELISA but also a list of the exact HCPs recognized by your ELISA antibodies. In addition, we provide access to our extensive database of HCPs of concern – based on current regulatory guidelines, literature, analyses of commercially released products, and our experience from more than 750 client projects – allowing data-driven risk assessment of the HCPs in your product. This is solid documentation for regulatory authorities.

Would you like to compare ELISAs from different vendors to select the best fit for your product?

Not all commercial HCP ELISAs may be equally suitable for detecting the specific HCPs in your biologic. Comparing the coverage of various ELISAs — commercial kits or process-specific assays — allows you to pick the best and most fitting ELISA for your drug and process-related impurities. Furthermore, you must ensure that the ELISA standard resembles the HCPs in your drug substance, or the readout may be off.

ELISA-MS™ combines ELISA-based immunocapture with LC-MS analysis to investigate which ELISA antibodies bind to your early process sample. The result is a very accurate and detailed list of individual HCPs in the sample recognized by the ELISA antibodies. You can then compare and base your choice of kit not only on the total coverage but also on each ELISA’s ability to detect individual HCPs, e.g. HCPs of concern. Furthermore, LC-MS can be used to analyze the fit of the ELISA standard to your drug substance.

On demand webinar

Genmab mAb HCP Control Strategy

How can your commercial HCP ELISA kit be approved for HCP analysis by the FDA and EMA?

We have invited Søren Skov Jensen and Margarita Sabater, Senior SMEs in Extended Characterization from the Late Stage Manufacturing Development group (CMC), to share Genmab's strategy for selecting the most appropriate release testing method, including selection of CHO anti-HCP ELISA reagents, identification of HCPs, and coverage analysis using ELISA-MS.

WATCH HERE

Senior CMC managers Søren Skov Jensen and Margarita Sabater from Genmab

Curious to know more about HCP coverage analysis?

Whatever ELISA-related challenge or question you may have, we are here to help you solve it. One of our protein analysis experts will discuss the best analysis approach or method for your project by email or online meeting – without obligation.

Typical project process

You typically work with
these experts:

Anette Holck Draborg

PhD in Immunology

Melissa Fernard Pedersen

MSc in Medicine with Industrial Specialization

Martine Mørck Taagholt

MSc in Biomedicine and industrial research experience

Project scope
We like to start with an online meeting to learn more about your project. Based on your needs, you will receive a draft proposal outlining the suggested analyses and expected timeframe.
Samples
After signing the final project proposal, we will contact you for details about your samples. An estimated report delivery date will be sent as soon as we receive your samples.
Execution
An Alphalyse project leader will oversee the project and send you status updates by email regularly.
A typical analysis includes:
- ELISA-MS™ analysis of selected HCP antibodies using a harvest sample.
- LC-MS-based HCP analysis of harvest, mock, and/or drug substance (DS) samples.
Results
You will receive the analysis report by email. Depending on the project, it includes:
- Objectives, description of analytical procedure, results, and conclusions.
- Reporting of HCPs covered by the ELISA antibodies in the sample and their distribution, total HCP coverage percentage.
- Evaluation and comparison of HCP profiles of the samples and/or to the ELISA standard.
- HCP ELISA potential high responder HCPs.
- HCP antibody binding of HCPs of potential concern (CHO projects only).
- Selected raw data.
Follow up
Upon project completion, your team is invited to go over the results at an online meeting.