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FabRICATOR® IdeS Antibody Cleavage

FabRICATOR® (IdeS)

FabRICATOR (IdeS) is a cysteine protease that digests antibodies at a specific site below the hinge, generating a homogenous pool of F(ab')2 and Fc/2 fragments after 30 min incubation.

 

FabRICATOR is available in seven different product formats:

  • FabRICATOR Enzyme - lyophilized enzyme
  • FragIT - immobilized on agarose beads
  • FragIT kit -  immobilized enzyme and affinity purification resin for generation and purification of F(ab')2 and Fc/2 fragments from IgG
  • FabRICATOR LE - lyophilized enzyme with low levels of endotoxins
  • FabRICATOR Validation kit - consists of three batches of the enzyme to simplify method validation
  • FabRICATOR-HPLC - immobilized enzyme in an HPLC column for fast online digestion of monoclonal antibodies
  • FabRICATOR MagIC - immobilized enzyme on magnetic agarose beads for fast parallel digestion of multiple antibodies

Also, a method to detect the FabRICATOR Enzyme is available:

  • Anti-FabRICATOR - a goat polyclonal antibody directed towards the FabRICATOR enzyme

Product Information

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, 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 expressed in E. coli. The enzyme contains a His-tag and has a molecular weight of 37 725 Da. FabRICATOR is a recombinantly modified form of IdeS that has been developed by Genovis AB for biotech applications requiring the highest standards and consistency.

  • 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)

Applications

Product Formats

FabRICATOR® Enzyme

FabRICATOR enzyme supplied as a lyophilized powder. The enzyme is available in 8 x 100 units, 96 x 100 units, 2000 units, 5000 units and 5 x 5000 units (1 unit digests 1 ug IgG).

FabRICATOR® LE

FabRICATOR enzyme lyophilized for endotoxin-sensitive assays. This product formulation has extremely low levels of endotoxins per vial (<0.04 EU /vial for the 2000 Unit vials). Available in 2000 and 5000 unit vials.

FabRICATOR® Validation Kit

The FabRICATOR Validation Kit contains three batches of the enzyme to simplify method validation. A certificate of analysis is included with each kit.

FragIT - Immobilized FabRICATOR®

FragIT is FabRICATOR enzyme immobilized on agarose beads, allowing for antibody subunit generation without contaminating the final preparation with FabRICATOR enzyme.

FragIT kit

FragIT Kit contains FragIT resin for antibody digestion and an affinity resin for purification of F(ab')2 and isolation of Fc fragments.

Anti-FabRICATOR™ Antibody

Anti-FabRICATOR is a goat polyclonal antibody directed towards the FabRICATOR enzyme.

FabRICATOR®-HPLC

FabRICATOR-HPLC is an immobilized enzyme in an HPLC column for fast online digestion of monoclonal antibodies.

  • A0-FR1-250

    FabRICATOR, 5×5000 units

    A0-FR1-2503,395.00Add to cart

The FabRICATOR Movie

 

FabRICATOR workflow

Middle down work flow using FabRICATOR enzyme and subsequent reduction. Sample preparation time approximately 2 hours.

Fabricator IdeS workflow

 

Download FabRICATOR posters

Brochures and Posters

Download Genovis brochures and posters with the latest details on the SmartEnzymes

SmartStories

FabRICATOR & GingisKHAN Assist in-source decay FT-ICR Mass Spectrometry analysis

FabRICATOR IdeS Applications

Applications

Learn more about the applications of Genovis SmartEnzymes

Download FabRICATOR Poster, Appnote and External References

Download FabRICATOR poster

ASMS 2013

"Optimizing the Enzymatic Subunit Generation with IdeS for High Throughput Structure Verification of Therapeutic Antibodies by Middle-Down Mass Spectrometry."

Download FabRICATOR poster

FabRICATOR Appnote

"Analyze the Fc glycosylation profile of a therapeutic antibody by using FabRICATOR digestion in combination with LC-MS."

 

FabRICATOR IdeS references

References

Find external articles and papers using FabRICATOR

 

 

 

FAQ and Support

Popular FAQ

FragIT digests rabbit IgG but to capture the Fc fragments in the affinity step you need another resin than the Capture Select Fc normally included in the kit. Please contact the Genovis team if you are interested in processing rabbit IgG and we will provide columns with resin compatible with rabbit Fc in the FragIT kit.

FabRICATOR MagIC is suitable for using in automated systems that can handle magnetic beads. On the applications page you can read about how it’s used in a KingFisher™ workflow.

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.

Yes, PNGaseF and FabRICATOR can be used in a one-pot reaction as long as the reaction conditions follow the specifications for the FabRICATOR enzyme. It is important to have native reaction conditions for FabRICATOR to remain active.

 

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