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FabRICATOR® Z Enzyme

FabRICATOR Z Enzyme is supplied as a lyophilized powder with no preservatives added, and is available in 2000 unit vials for digestion of up to 2 mg mouse IgG2a and IgG3.

 

FabRICATOR Z (IdeZ) is a cysteine protease that digests mouse IgG2a and IgG3 at a specific site below the hinge, generating a homogenous pool of F(ab')2 and Fc fragments after 2 hours incubation.

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Detailed Information

FabRICATOR Z digests mouse IgG2a and IgG3 at a specific site below the hinge region, producing a homogenous pool of F(ab')2 and Fc fragments. Most mouse IgG2a are digested by FabRICATOR Z within 2 hours although the enzyme:ab ratio and/or digestion time may need to be optimized for certain antibodies. There is no risk of overdigestion if the incubation time is prolonged. Since FabRICATOR Z digests IgG under physiological reaction conditions, the immunoreactivity is preserved.

FabRICATOR Z is derived from S. equi ssp. zooepidemicus and expressed in E. coli. The enzyme contains a His-tag and the molecular weight is 36 442 Da.

Storage and Stability

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

Unit Definition

One unit digests ≥ 95% of 1µg mouse IgG2a when incubated in 10 mM Tris, 10 mM NaCl, pH 6.5 at 37°C for 2 hours.

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

Instructions

Product Specification

Safety Data Sheet

FAQ and Support

Latest FAQ

There should not be any problems to use low concentrations of IgG. If you don't get complete digestion, try to increase the incubation time. There is no risk of over digestion and the incubation time can be prolonged to over night if necessary.
An alternative would be to increase the ratio enzyme to antibody (i.e. Follow protocol for mouse IgG). We have not made any specific tests on low concentrations but we have not heard that it would be any problems.

Yes, FabRICATOR can cleave Fc-Fusion proteins. For example Enbrel is effectivelty cleaved by FabRICATOR into Fc and TNFR. Enbrel (etanercept) is a fusion protein composed of the Fc domain of hIgG1 and the p75 tumor necrosis factor receptor (TNFR).

FabRICATOR is very specific and small changes in the sequence (..CPAPELLGGPSVF..) may reduce or completely impair the ability to digest the IgG.

We have tested a few different modified sequences and we can see that the enzyme digest the IgG to some degree to Fab2 and Fc under somewhat modified reaction conditions. We then obtained a mixture of Fc, Fab2 and single cut mAb (one of the HC still undigested).

We used PBS buffer but with greatly reduced salt concentration (5-10 mM instead of 140 mM) which appeared to promote digestion. We also added 10X more enzyme than recommended amount and prolonged incubation time.

It is very difficult to predict the outcome for a specific modification. It might work, but we cannot guarantee it unfortunately.

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Patent and Disclaimer


FabRICATOR®


Genovis AB hold a license for exclusive worldwide use of biotechnical industrial applications of IdeS covered by any patents derived from International publication WO03051914, including granted US Patent No US7666582.
Other aspects of FabRICATOR®technology are encompassed by pending patent applications in the name of Genovis AB.

The trademark FabRICATOR® is the property of Genovis AB.


For research use only. Not intended for any animal or human therapeutic diagnostic use.

©2018 Genovis AB.