HCP analysis of monoclonal antibodies
- Check for HCPs that are not detected by ELISA
- Investigate if your mAb contains HCPs known to be problematic
- Compare individual HCPs in a biosimilar against the originator
Do you want to check your mAb for problematic HCPs?
Monoclonal antibodies are generally very clean due to efficient protein A purifications. However, it is still necessary to document the HCP level to the authorities.
Typically, you also want to confirm the HCP ELISA results by an orthogonal LC-MS analysis and check for problematic HCPs. For instance HCPs that degrade the drug product, e.g. phospholipase a...
Do you want to check your mAb for problematic HCPs?
Monoclonal antibodies are generally very clean due to efficient protein A purifications. However, it is still necessary to document the HCP level to the authorities.
Typically, you also want to confirm the HCP ELISA results by an orthogonal LC-MS analysis and check for problematic HCPs. For instance HCPs that degrade the drug product, e.g. phospholipase and cathepsin, or HCPs that are immunogenic in patients, e.g. clusterin.
Lower your limit of detection and check for problematic HCPs
Often, HCP ELISAs have sufficient coverage of HCPs from the CHO expression system. However, ELISAs have poor specificity and provide only a number for the total HCP level. Thus, abundant HCPs and potentially problematic HCPs can go undetected by ELISA. Especially Host Cell Proteins of low MW and highly conserved proteins, e.g. ubiquitin, are poorly immunogenic. As a result, they are easily missed by the ELISA.
However, there is a good solution to this problem:
By using our best practice LC-MS method you get the total HCP amount by an orthogonal method to ELISA. In addition, you get a list of the individual HCPs and the amount of each. Moreover, by combining with a native digest which is optimized for mAbs, we can even lower the limit of detection to sub-ppm levels.
Check for specific Host Cell Proteins
As a unique service, our specialists will both check for specific HCPs that are known to be of concern in mAb products, and also compare to HCPs found in our study of commercially available mAbs. You can then use this info to ensure the stability and efficacy of your drug. Similarly, you can make changes to lower the risk of immunogenic reactions in patients.
What is included in an orthogonal LC-MS assay:
Our LC-MS method was developed and tested on hundreds of mAb samples, including 16 commercial drugs. It is also worth noting that an assay for your mAb can be set up in a matter of weeks.
Analysis of HCP in mAbs includes the following:
- An optimized sample preparation, specifically developed for monoclonal antibody samples.
- Analysis on a robust microflow HPLC and SWATH® MS/MS system to ensure high reproducibility.
- Spike-in of internal standard proteins (0-2000 ppm). This step is important since it enables quantification linearity even to low ppm levels.
- A report with both the total HCP number and a list of individual HCPs with amounts.
- A check for potentially problematic HCPs, and a check for HCPs found in commercial products.
- And finally, a follow-up meeting to go over the results.
