FabRICATOR®
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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.
FabRICATOR Lyophilized is available as a lyophilized powder in 2000 (A0-FR1-020), 5000 (A0-FR1-050) and 5 x 5000 (A0-FR1-250, value pack) unit vials for digestion of 2, 5 and 5 x 5 mg IgG respectively, and in 8 x 100 unit strip wells (A0-FR1-008) and 96 x 100 unit well plates (A0-FR1-096) for digestion of 8 x 100 µg and 96 x 100 µg IgG respectively.
Why FabRICATOR® Lyophilized?
- Flexible format for method development
- Possible to combine enzymes for improved efficiency
- Amenable to automated workflows
- Ready to use – just add water!
Available Products
FabRICATOR® Lyophilized 2000 units
Lyophilized enzyme for below hinge digestion of 2 mg IgG
FabRICATOR® Lyophilized 5000 units
Lyophilized enzyme for below hinge digestion of 5 mg IgG
FabRICATOR® Lyophilized 5 x 5000 units
Lyophilized enzyme for below hinge digestion of 5 x 5 mg IgG
FabRICATOR® Lyophilized 8 x 100 units
Lyophilized enzyme for below hinge digestion of 8 x 100 𝜇g IgG
FabRICATOR® Lyophilized 96 x 100 units
Lyophilized enzyme for below hinge digestion of 96 x 100 𝜇g IgG
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Product Information
Unit Defintion
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.
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
Content and Storage
FabRICATOR Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, the enzyme 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.
Resources
Instruction for Use
Product Specifications
Safety Data Sheet
Certificate of Analysis
Find your certificate of analysis »AppNote SmartEnzymes™ for mAb Oxidation
Download Application Notes
AppNote FabRICATOR® for Glycan Analysis
Download Application Notes
SmartEnzymes™ for Generation of Homogeneous Antibody Fragments
Download Scientific Poster
How FabRICATOR® works
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Applications
Fast and reliable middle-level analysis of antibody oxidation using FabRICATOR and GlycINATOR, supporting robust QC of critical quality attributes.
Performance Activity
Generation of defined IgG subunits via single-site, below hinge cdigestion, enabling high-resolution middle-level LC-MS and improved PTM characterization.
Citations
Powered by BiozFAQ and Support
Can I re-use FabRICATOR Z Immobilized columns?
We can only guarantee optimal digestion for one-time use. We do not have a cleaning or regeneration protocol to provide. However, depending on the antibody and the following application the column can be reused. We do not recommend using with different antibodies due to the risk for contamination of carry-over from previous sample. If column reuse is desired store the column in 10-20% ethanol at +4-8°C.
Is it possible to use FabRICATOR in a buffer containing EDTA or HSA?
Yes. FabRICATOR is compatible with EDTA. Make sure your pH is within 6-8.
Does FabRICATOR introduce modifications?
FabRICATOR is a cysteine protease. It should not result in any other modifications than the digestion of the peptide bond. Digestion will be between the two Glycines (…CPAPELLG / GPSVF…) and the C-terminus will regain the carboxyl group – that is addition of one OH-group and the N-terminal amino group will be regained (addition of one H).
In summary, R1-CO-NH-R2 + H2O –> R1-COOH + NH2-R2.
Can I use FabRICATOR (IdeS) with very low amounts of IgG?
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.
Which digestion buffers are compatible with FabRICATOR HPLC?
We have tested PBS and 150 mM ammonium acetate, pH7 with good results. More buffers were found to be compatible with FabRICATOR digestion in solution (more info) and should be suitable for FabRICATOR HPLC as well. However, the ionic strength of the buffer influences retention of the antibody fragments on the FabRICATOR HPLC column and a minimum of 100-150 mM salt is necessary for IgG1 subunits to elute as a single, narrow peak. Other IgG subclasses or fusion proteins might need some optimization of buffer composition.
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