SmartEnzymes 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.

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
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)

FabRICATOR® Lyophilized

Lyophilized enzyme for below hinge digestion of IgG

FabRICATOR® Immobilized

Immobilized enzyme for below hinge digestion of IgG in spin columns

FabRICATOR® Fab2 Kit

Immobilized enzyme and affinity resin for below hinge digestion of IgG and purification of fragments

FabRICATOR® HPLC

Immobilized enzyme for on-column digestion of IgG below the hinge

FabRICATOR® MagIC

Immobilized enzyme on magnetic beads for automated digestion of IgG below the hinge

FabRICATOR® Low Endotoxin

Lyophilized enzyme with low levels of endotoxin for below hinge digestion of IgG

FabRICATOR® Validation Kit

Three different lyophilized batches for validation of FabRICATOR-based analytical methods

About FabRICATOR® (IdeS)

FabRICATOR workflow

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

FabRICATOR® (IdeS) Citations

FAQ and Support

Popular FAQ

The protocol for FabRICATOR MagIC is based on using standard PBS pH 7.4 as digestion buffer. FabRICATOR in solution is compatible with most commonly used buffers at pH=6-8 and the same buffers should work for FabRICATOR MagIC, but the incubation time might need to be optimized.

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.

The FabRICATOR MagIC magnetic beads with immobilized FabRICATOR (IdeS) enzyme.

The FabRICATOR HPLC can run >400 samples of 10 ug of IgG.

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.

 

Find all FAQ here

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