FabRICATOR

FabRICATOR® Enzyme

The FabRICATOR Enzyme is supplied as a lyophilized powder with no preservatives added, and is available in 2000 and 5000 unit vials for digestion of up to 2 - 5 mg IgG.

 

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.

Detailed 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. There is no other known substrate besides IgG. 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.

Digestion of IgG using FabRICATOR followed by reduction, produces three 25 kDa-sized fragments (Fd’, Fc and LC) that can be easily analyzed using LC-MS instrumentation. The high-accuracy mass profile enables detection of antibody microvariants such as glycosylation, C-terminal lysine clipping, oxidation and deamidation and several others.

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.

Storage and Stability

The FabRICATOR enzyme is supplied lyophilized and reconstituted by addition of water. After reconstitution, FabRICATOR 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.

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

Unit Definition

One unit digests ≥ 95% of 1µg human IgG when incubated in 10 mM sodium phosphate, 137 mM NaCl, 2.7mM KCl pH 7.4 at 37°C for 30min.

  • A0-FR1-096

    FabRICATOR, 96×100 Units

    A0-FR1-0961,735.00Add to cart
  • A0-FR1-250

    FabRICATOR, 5×5000 units

    A0-FR1-2503,395.00Add to cart

How to Order

For details on ordering please visit How to order. Click add to cart to buy online.

Quotations

Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities.

Documents

FAQ and Support

Popular FAQ

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.

Yes, for human IgG1 molecules both enzymes can be used in combination to generate a specific hinge peptide. The incubation time is significantly longer for FabALACTICA and it is recommended to first run the FabALACTICA digest and then add FabRICATOR for the last 30 min.

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.

Yes. FabRICATOR is compatible with EDTA. Make sure your pH is within 6-8.

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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Legal and Disclaimers

All rights reserved. Genovis products are covered by one or more patents, pending patent applications, trademarks and/or copyrights owned or controlled by Genovis AB.

For more information about commercial rights, please contact Genovis team at info@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.

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