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-FR4-0601,550.00Add to cart

How to Order

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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

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

No. According to the following reference, FabRICATOR does not digest IgM. Pavel-Rammingen et al., 2002. IdeS, a novel streptococcal cysteine proteinase with unique specificity for immunoglobulin G.

FabRICATOR is a cysteine protease. It should not result in any other modifications than the digestion of the peptide bond. Digestion will be between the two Glycines (…CPAPELLG / GPSVF…) and the C-terminus will regain the carboxyl group – that is addition of one OH-group and the N-terminal amino group will be regained (addition of one H).

In summary, R1-CO-NH-R2 + H2O –> R1-COOH + NH2-R2.

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.

 

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