Services Data for risk assessment

Data for risk assessment

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

Regulatory authorities are increasingly aware that protein impurities, even at low levels, can affect patient safety and product stability. Some residual Host Cell Proteins (HCPs) have been observed to cause immunogenicity, adjuvant effects, or direct biological activity in patients and degradation of products or excipients during storage. Therefore, it is not enough that an ELISA reports a low number – you also need to investigate the individual HCPs.

For this reason, legal authorities call for an orthogonal method, such as mass spectrometry (LC-MS analysis), to identify and quantify individual HCP impurities and for a thorough risk assessment of the resulting data. LC-MS can identify process-related impurities down to 0.1 ppm and allows for data-driven process optimization and evaluation of purification steps.

We use relevant ICH guidelines, protein standards, and follow the US Pharmacopeia General Chapter <1132.1>, resulting in very robust and comparable results, delivered in just 2-4 weeks.

Why use LC-MS/MS for HCP analysis?

Patient safety and risk assessment

Rule out the presence of HCPs causing immunogenicity, adjuvant effects, or biological activity

Product stability and efficacy

Check for enzymes causing product degradation, modification, loss of activity, or excipient degradation

Manufacturing consistency and product purity

Evaluate purification process clearance, drug substance purity, or batch comparability

Why should you use an orthogonal method – if ELISA shows that your product is pure?

HCP-ELISAs tend to be biased towards abundant HCPs and are inherently immunologically weighted, which means they may fail to detect trace or non-immunogenic impurities in the final product. In addition, the readout of immunoassays is an aggregate sum of present HCPs, which does not confirm or deny the presence of specific HCPs of concern.

Using mass spectrometry (MS), you will know which specific HCPs your product contains – and can evaluate if any of them may be of concern.

Do you suspect that enzyme impurities affect your product's stability or efficacy?

Active enzymes need only be present at shallow levels to cause product or excipient degradation, or modification. This can lead to loss of drug activity (efficacy) or drug product stability upon storage. Examples of enzymes with significant impact on product or excipient stability include proteases which cleaves the protein biopharmaceutical, and tween-degrading enzymes; lipases, and esterases, which degrade Polysorbate 20 and Polysorbate 80 used as solubilizing excipient in the formulation buffer of many mAbs and biopharmaceuticals.

ELISAs are in general not able to detect specific HCPs at low ppm levels. By relying on HCP-ELISA only, you run the risk that your product may contain one or more HCPs of concern, which may degrade your product or cause adverse reactions in patients.

Do you know how impurities in your biosimilar differ from the originator?

Since biosimilar products can be produced in different expression systems and are likely purified using processes other than the originator, the HCP impurity profile in the two products may differ.

Authorities ask that biosimilars are subject to just as extensive HCP analysis as their originators to verify that they do not contain HCPs of concern. In some cases, we have found that a biosimilar product is even cleaner than the originator, which has served as a quality attribute of the product.

On demand webinar

Which HCPs to look for and how to measure them

In several clinical trials and during the development of many biologics, residual HCPs, such as flagellin, proteases, and the infamous PLBL2, have proved problematic. They may cause immunogenicity in patients, unwanted biological effects, or enzymatically degrade the drug, decreasing its efficacy and shelf life.

For mAbs, it is now well-described which HCPs from the CHO expression system to be concerned about. Regulatory guidelines assert that documentation should include risk assessments of the individual HCPs identified and removed.

Problematic HCPs can stop your biologics program!

Typical project process

You typically work with
these experts:

Melissa Fernard Pedersen

MSc in Medicine with Industrial Specialization

Peter Toft Jensen

PhD in Biochemistry and Molecular Biology

Dean Malte Zisco

MSc in Biochemistry and Molecular Biology

Freja Andersen

MSc in Biomedicine

Project scope
We will 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 at regular intervals.
A typical analysis includes:
- Spike-in of internal standard proteins, enabling quantification at low ppm levels.
- Enzymatic digest and analysis of peptides using LC-MS.
- ELISA-MS analyses with an in-well digest of immunocaptured HCPs.
- Setup of a custom-specific database.
Results
You will receive the report by email. As standard, the LC-MS HCP analysis report includes:
- Objectives, description of analytical procedure, results, and conclusions.
- A list of the number and the total amount of HCPs in the sample.
- A list of individual HCPs and their quantities, highlighting HCPs of potential concern, including the physiochemical properties of the HCPs.
- The coverage percentage of HCPs in ELISA.
- Match between the ELISA standard and the harvest or mock samples.
- ELISA Coverage of HCPs found in DS, including coverage of potential problematic HCPs.
- Selected raw data, e.g., excel sheets.
Follow up
Upon completion of the project, your team is invited to go over the results at an online meeting.

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Curious to know more?

Whatever 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.

Client stories

Accelerating early-stage process development: eleva case study

Detecting and quantifying HCPs in a moss-based system

Data for risk assessment

Why use LC-MS/MS for HCP analysis?

Patient safety and risk assessment

Product stability and efficacy

Manufacturing consistency and product purity

Why should you use an orthogonal method – if ELISA shows that your product is pure?

Do you suspect that enzyme impurities affect your product's stability or efficacy?

Do you know how impurities in your biosimilar differ from the originator?

On demand webinar

Typical project process

Project scope

Samples

Execution

Results

Follow up

Curious to know more?

Client stories

Benefits of LC-MS HCP analysis for risk assessment

What clients say

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