Ask The Expert - Verifying Removal of Residual Production Enzymes from Your Final Product

Published: June 13, 2016
Session Closed
Session Expert: Dr. Roger Noble , Roche Custom Biotech

Enzymes such as Trypsin and Collagenase are often critical components for product processing, modification of proteins and cell preparation for stem cell therapies. As these final cell-based products are intended for administration into patients, it is important that any residual enzyme material from the production process be assessed and its final concentration minimized to reduce risk. Potential negative impacts of residual enzymes could include immunogenic activity, reduced efficacy of the final product, and toxicity in patients. Although there are no specific regulatory guidelines at this time to specify allowable amounts of residual Trypsin or Collagenase, these are still considered process residuals and potential product contaminants, and thus should be assessed as part of an overall risk mitigation strategy.

While activity assays are often used to verify removal of residual enzymes, these have limited sensitivity and may only provide partial information as they will only detect active enzyme, while missing inactive fragments that might still be immunogenic. ELISA technology that is independent from activity is a favorable alternative for detecting residual enzymes such as Trypsin and Collagenase.

Questions and Answers

At what stage(s) of the production cycle should you test for residual enzymes?

This is very much up to the discretion of the individual company and your risk mitigation strategy. Testing can be done as an in-process control to monitor efficiency of the… Read More

What type of samples are compatible with your assays? Is there any sample preparation required?

The assays are intended for use with aqueous buffer solutions from biotechnology processes and cell culture supernatant. If cell-free aqueous solutions are used, no sample preparation is required. For testing… Read More

Do you have a clinical grade trypsin?

Roche’s Recombinant Trypsin is manufactured under ISO 13485 and in accordance to good manufacturing practice (GMP) guidelines. Additionally, no animal derived products are used in the fermentation, purification, or final… Read More

We use BAPNA to test for residual trypsin activity. How does your kit compare?

The Residual Protein Trypsin kit is an antibody-based immunoassay which provides data indicating the quantity of Trypsin protein present in a sample. The BAPNA test detects the rate of enzyme… Read More

We are interested in conducting in process testing for residual enzymes to check our process efficiency at a couple stages. Can you explain what kind of information we will gain by running your kits particularly with respect to in process tests? Also do you have any recommendations for using you kits in this manner?

These ELISA kits are intended for quantitative detection of the respective enzyme proteins in manufacturing processes of biopharmaceutical and cell therapy products. They can be used with sample material from… Read More

What if you are not using Roche enzymes, can you still use these kits?

The Residual Protein Liberase Kit was designed to detect the constituents of Liberase Blends containing Collagenase I, Collagenase II, and Thermolysin, specifically the formulations found in the following Liberase Blends… Read More

What is the time to results for these assays?

Total assay time is approximately 2 hours and 30 minutes. Incubation time is 1 hour and 55 minutes. Therefore, minimal hand-on time is required to run these assays. Read More

What are the sensitivity and measuring ranges of your assays?

The Residual Protein Trypsin assay has a sensitivity of ≤0.5 ng/ml and measuring range of 0.5 ng/ml up to approximately 50 ng/ml. The Residual Protein Liberase kit has a sensitivity… Read More

Is using your kits easier than preparing homebrew ELISA assays?

The Roche ELISA assays can be performed in any laboratory without specialized training. Additionally, they are available in an all-inclusive kit format with validated reagents, so all components necessary are… Read More

What type of enzymes will be detectable with these assays?

The Residual Protein Trypsin assay detects Roche Recombinant Trypsin from porcine pancreas, expressed in Pichia Pastoris. The Residual Protein Liberase assay detects constituents of the Roche Liberase Blends containing Collagenase… Read More