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, 5000 and 5 x 5000 (value pack) unit vials for digestion of 2, 5 and 5 x 5 mg IgG respectively, and in 8 x 100 unit strip wells and 96 x 100 unit well plates 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
Watch video
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
How much of the FabRICATOR do you need to cleave IgG in 50 µl of normal human serum?
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.
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.
Does FabRICATOR (IdeS) have protease activity on IgM?
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.
Can I cleave Fc-fusion proteins with FabRICATOR?
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.
Where does FabRICATOR cleave the IgG molecule?
FabRICATOR® cleave the IgG molecule in one specific site in the lower hinge region of the anitbody leaving a F(ab’)2 fragment and a Fc-fragment.
Human IgG1: ..CPAPELLG / GPSVF..
Human IgG2: ..CPAPPVA / GPSVF..
Human IgG3: ..CPAPELLG / GPSVF..
Human IgG4: ..CPAPEFLG / GPSVF..
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