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
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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
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..
I have a small modification in the sequence below the hinge, will FabRICATOR still digest my IgG?
FabRICATOR is a very specific enzyme and small changes in the sequence in or around the digestion site (…CPAPELLG / GPSVF…) may reduce or completely impair the ability to digest the IgG. It is very difficult to predict the outcome for each specific modification. It might work, but we cannot guarantee it unfortunately, the only way of knowing is to perform a digestion test. Some suggestions that are worth testing if you experience incomplete digestion: Increase the incubation time, increase the amount of added enzyme, use a PBS with greatly reduced salt concentration (5-10 mM instead of 140 mM) and pH 6.5 instead of 7.4.
Can FabRICATOR be stored in frozen stock solutions?
We have no stability data for storage at -20°C or -70°C. However, we have customer reporting that they successfully stored reconstituted FabRICATOR at -20°C or -70°C for a couple of months. Genovis cannot guarantee the activity of the enzyme, but these storage conditions might be OK depending on the intended application.
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.
Is FabRICATOR stable in the presence of denaturing agents such as guanidine hydrochloride or SDS?
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.
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