FabRICATOR

FabRICATOR® Validation Kit

The FabRICATOR enzyme has become a widely used tool in the biopharmaceutical industry for characterizing monoclonal antibodies, Fc-fusion proteins, ADCs and biosimilars for therapeutic use. Each batch of the FabRICATOR enzyme is rigorously tested for enzymatic activity, identity and purity before it is released. The FabRICATOR Validation Kit consists of three lots of the enzyme, to make the method validation easier. A certificate of analysis is included with each kit.

 

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. 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.

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 a recombinantly modified form of IdeS from Streptocoocus pyogenes that has been developed by Genovis AB for biotech applications requiring highest standard and consistency.

Digestion of IgG using FabRICATOR followed by reduction, produces three 25 kDa-sized fragments (Fd’, Fc and LC) that easily can be 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.

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-FR4-060

    FabRICATOR Validation Kit

    A0-FR4-0601,420.00Add 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

Product Specifications

Safety Data Sheet

Certificate of Analysis

FAQ and Support

Popular FAQ

FabRICATOR® cleave the IgG molecule in one specific site in the lower hinge region of the anitbody leaving a F(ab’)2 fragment and a Fc-fragment.
Human IgG1: HTCPPCPAPELLG / GPSVF
Human IgG2: VECPPCPAPP_VA / GPSVF
Human IgG3: PPCPRCPAPELLG / GPSVF
Human IgG4: PHAHHAQAPEFLG / GPSVF

FabRICATOR is a very specific enzyme and small changes in the sequence in or around the digestion site (…CPAPELLG / GPSVF…) may reduce or completely impair the ability to digest the IgG. It is very difficult to predict the outcome for each specific modification. It might work, but we cannot guarantee it unfortunately, the only way of knowing is to perform a digestion test. Some suggestions that are worth testing if you experience incomplete digestion: Increase the incubation time, increase the amount of added enzyme, use a PBS with greatly reduced salt concentration (5-10 mM instead of 140 mM) and pH 6.5 instead of 7.4.

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, 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.

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.

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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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