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 enzyme removal procedure, as well as for a final release testing criteria. The Roche assays offer a broad measuring range…

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 of cell culture samples, the samples must be centrifuged and the supernatants can be used for enzyme determination.

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 formulation. Roche’s Recombinant Trypsin is currently used as a critical raw material for the production of active pharmaceutical ingredients (API),…

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 activity in the sample, i.e. enzymatic cleavage of the BAPNA molecule. In other words, the BAPNA test indicates how much…

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 aqueous buffer solutions from biotechnological processes, as well as cell culture supernatant. The wide measuring ranges for each assay make…

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 available from Roche: Liberase MNP-S, Liberase MNP-S GMP Grade, and Liberase MTF C/T GMP Grade. The Residual Protein Trypsin Kit…

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.

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 of 0.1 ng/ml and measuring range of 0.1 ng/ml up to approximately 5 ng/ml.