Powered by BiozBelow 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.
The enzyme 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.
- 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
FabRICATOR® Lyophilized
Lyophilized enzyme for below hinge digestion of IgGFabRICATOR® Immobilized
Immobilized enzyme for below hinge digestion of IgG in spin columnsFabRICATOR® Fab2 Kit
Immobilized enzyme and affinity resin for below hinge digestion of IgG and purification of fragmentsFabRICATOR® HPLC
Immobilized enzyme for on-column digestion of IgG below the hingeFabRICATOR® MagIC
Immobilized enzyme on magnetic beads for automated digestion of IgG below the hingeFabRICATOR® Low Endotoxin
Lyophilized enzyme with low levels of endotoxin for below hinge digestion of IgGFabRICATOR® Validation Kit
Three different lyophilized batches for validation of FabRICATOR-based analytical methodsAbout FabRICATOR® (IdeS)
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
Oxidation Analysis
Fc-glycan Analysis
ADC Characterization
Subunit Analysis
Generate Antibody Subunits
Automated Subunit Analysis
Automated Parallel Analysis of AntibodiesFabRICATOR® (IdeS) Citations
FAQ and Support
Popular FAQ
FabRICATOR HPLC is an HPLC column with the FabRICATOR (IdeS) protease immobilized on a HPLC compatible matrix for automated antibody subunit generation.
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.
Yes, it is possible to digest the antibody in the first dimension and then separate the subunits in the second dimension. Please see the application note for detailed instructions.
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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