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FabRICATOR® – Below Hinge Digestion of IgG (IdeS)
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.
SmartEnzymes™
FabRICATOR is widely used in characterization, quality control, stability testing, production monitoring and clone selection of antibody-based therapeutics such as mAbs, ADCs, biosimilars and Fc-fusion proteins.
Why FabRICATOR®?
- Highly specific – one digestion site below the hinge
- Fast – generates a homogenous pool of fragments within 30 minutes
- Enables characterization and quality control of antibody-based biotherapeutics
- Available immobilized in ready-to-use spin columns
Human IgG1-4, IgG of some classes from monkey, rat, rabbit and sheep
30 min reaction
No need for reducing agents or co-factors
CPAPELLG / GPSVF (below the hinge)
Product Formats
FabRICATOR® Immobilized
Immobilized enzyme for below hinge digestion of IgG in spin columns
Buy productFabRICATOR® Fab2 Kit
Immobilized enzyme and affinity resin for below hinge digestion of IgG and purification of fragments
Buy productFabRICATOR® MagIC
Immobilized enzyme on magnetic beads for automated digestion of IgG below the hinge
Buy productFabRICATOR® Low Endotoxin
Lyophilized enzyme with low levels of endotoxin for below hinge digestion of IgG
Buy productFabRICATOR® Validation Kit
Three different lyophilized batches for validation of FabRICATOR-based analytical methods
Buy product
About FabRICATOR®
FabRICATOR (IdeS) is a unique enzyme that digests IgG at a specific site below the hinge region, generating a homogenous pool of F(ab’)2 and Fc/2 fragments.
No other substrate besides IgG is known. The IgG is digested within 30 minutes, and there is no risk of overdigestion if the incubation time is prolonged. Since FabRICATOR digests IgG under physiological reaction conditions, the immunoreactivity is preserved. The best activity is obtained at 37°C, but digestion can also be performed at room temperature using a prolonged reaction time.
FabRICATOR digests all subclasses of human and some classes of monkey, rabbit and sheep IgG. It has limited activity on mouse IgG2a and IgG3 – for digestion of these antibody species, we recommend using FabRICATOR Z.
FabRICATOR is cloned from Streptococcus pyogenes and recombinantly expressed in E. coli. It is produced by Genovis AB for analytical and biotech applications requiring the highest standards and consistency. The enzyme contains a His-tag and has a molecular weight of 38 kDa.
A method to detect the FabRICATOR enzyme is available. Learn more about Anti-FabRICATOR, a goat polyclonal antibody.
Applications
Fast and reliable middle-level analysis of antibody oxidation using FabRICATOR and GlycINATOR, supporting robust QC of critical quality attributes.
Performance Activity
Generation and purification of F(ab’)2 and scFc fragments without residual enzyme, enabling rapid preparation of antibody fragments for downstream applications.
Generation of defined IgG subunits via single-site, below hinge cdigestion, enabling high-resolution middle-level LC-MS and improved PTM characterization.
Citations
Resources
How FabRICATOR® works
Watch video
SmartEnzymes™ for Automated and High-throughput Characterization of Antibodies
Download Scientific Poster
AppNote FabRICATOR® MagIC for Automated Analysis in Complex Samples
Download Application Notes
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
SmartStories
FabRICATOR® MagIC produce F(ab)2 subunits in less than 30 minutes
Dan Bach Kristensen
Symphogen A/S
FabRICATOR® Supports Analysis of a Complex Antibody-Oligonucleotide Conjugate
Yury Tsybin
Spectroswiss
FabRICATOR® for Intact Analysis of Biologics
Min Kyung So
KBIO Osong Medical Innovation Foundation
FAQ and Support
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.
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 use protein A or MabSelectSure (GE) for removal of Fc-fragments?
Genovis has chosen CaptureSelect in the FabRICATOR Fab2 Kit for removal of Fc fragments. CaptureSelect has a very high specific affinity for the Fc. Both protein A and MabSelect Sure (GE) may have affinity for the Fab which may result in a lower recovery of the generated F(ab’)2 fragments and the degree of affinity is antibody dependent. We have many customer reports on this subject, and they usually come back to us for the CaptureSelect columns for the above reason.
Can I use protein A or MabSelectSure (GE) for removal of Fc-fragments?
Genovis has chosen CaptureSelect in the FabRICATOR Fab2 Kit for removal of Fc fragments. CaptureSelect has a very high specific affinity for the Fc. Both protein A and MabSelect Sure (GE) may have affinity for the Fab which may result in a lower recovery of the generated F(ab’)2 fragments and the degree of affinity is antibody dependent. We have many customer reports on this subject, and they usually come back to us for the CaptureSelect columns for the above reason.
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.
Legal & Disclaimers
All rights reserved. Genovis products may be covered by one or more patents, trademarks and copyrights owned by Genovis AB or licensed from third parties. For more information about commercial rights, please contact the Genovis team at licensing@genovis.com.
Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals. All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.
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