Validation Kit
Three different lyophilized batches for validation of FabRICATOR-based analytical methods. FabRICATOR (IdeS) is an IgG-specific cysteine protease that digests antibodies at a single amino acid site below the hinge, generating homogenous F(ab’)2 and Fc fragments.
The FabRICATOR enzyme has become a widely used tool in the biopharmaceutical industry for characterizing monoclonal antibodies, Fc-fusion proteins, ADCs and biosimilars for therapeutic use.Each batch of the FabRICATOR enzyme is rigorously tested for enzymatic activity, identity and purity before it is released. The FabRICATOR Validation Kit consists of three batches of the lyophilized enzyme, to make the method validation easier. A certificate of analysis is included with each batch.
- Simplified method validation for QC applications
- Ideal format for batch-to-batch variability assessment
- Increased confidence in performance of batch release assays
Available Products
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Download product folderUnit Definition
One unit digests ≥ 95% of 1µg human IgG when incubated in 10 mM sodium phosphate, 137 mM NaCl, 2.7 mM KCl pH 7.4 at 37°C for 30 min.
Storage and Stability
FabRICATOR Validation Kit consists of three different lyophilized batches of the FabRICATOR Lyophilized enzyme. The enzymes are reconstituted by addition of water. No preservatives added. After reconstitution, FabRICATOR is stable for 1 month at +4-8°C. The product is shipped at ambient temperature, and should be stored at -20°C upon arrival.
Buffers
The buffers below have been tested for compatibility with FabRICATOR digestion at different pH. Note that the FabRICATOR enzyme is irreversibly inactivated below pH 5.0. Digestion above pH 8.0 requires prolonged/optimized conditions.
- Phosphate buffer saline (PBS) 6.0 – 8.0
- Tris buffer 7.0 – 8.0
- MES buffer 5.5 – 6.5
- HEPES buffer 7.0 – 8.0
- Ammonium bicarbonate buffer 6.0 – 7.0
- Sodium acetate buffer 6.0
Instructions
Product Specifications
Safety Data Sheet
Certificate of Analysis
FAQ and Support
Popular FAQ
If we assume that the antibody concentration in serum is 15 mg/ml you would have to add 10-15 000 units to a sample of 1 ml serum. In 50 µl serum you would have to add 500 – 750 units of FabRICATOR.
It is possible to use FabRICATOR with low concentrations of IgG. Low concentrations of IgG will lead to decreased reaction rate, so for concentrations below 0.5mg/mL it is therefore recommended to increase the amount of added enzyme and/or increase the incubation time. There is no risk of over-digestion and the incubation time can be prolonged to O/N if necessary.
We have tested 4-6 M GuHCl which the enzyme does not withstand without significant decrease of activity. SDS have not been evaluated but 0.1% RapiGest from waters have negative influence on activity. Taken together the enzyme does not well tolerate denaturing agents although we have not tested this in more detailed studies.
No. According to the following reference, FabRICATOR does not digest IgM. Pavel-Rammingen et al., 2002. IdeS, a novel streptococcal cysteine proteinase with unique specificity for immunoglobulin G.
Yes, FabRICATOR should be able to digest a Fc fusion protein as long as you have not fused the protein too close to the cleavage site. FabRICATOR digests human IgG just below the hinge (…CPAPELLG / GPSVF…), leaving the hinge on the fusion-part.
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