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FabRICATOR Z Enzyme

FabRICATOR Z is a recombinantely modified IdeZ protease from Streptococcus equi ssp. zooepidemicus digests mouse IgG2a and IgG3 and produces a homogenous pool of F(ab')2 and Fc fragments after 2 hours incubation.

  • More effective than FabRICATOR (IdeS) on mouse IgG2a
  • Works at neutral pH
  • Safe - no risk of over digestion

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Products

FabRICATOR Z enzyme is supplied as lyophilized powder with no preservatives added and is available in 2000 unit pack size.

Quotations

Don't hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities.

Detailed Information 


FabRICATOR Z digests mouse IgG2a and IgG3 at one 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.

Since FabRICATOR Z only cleaves at one specific site, below the hinge region, there is no risk of over digestion if the incubation time is prolonged for several hours. FabRICATOR Z digests IgG under physiological reaction conditions thus preserving the immunoreactivity.

Digestion Site

The primary digestion site in mouse IgG2a is ...CPAPNLLG / GPSVF...

Follow this link to view a protease IgG digestion chart with species 

Stability

The enzyme is reconstituted by addition of water and after reconstitution FabRICATOR Z is stable for 1 month at +4-8°C. The product is shipped at ambient temperature but 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.

Documents

Instructions

Product specifications

MSDS

Enzyme Reference Tool

FabRICATOR is used frequently for studying important quality attributes of antibody-based biotherapeutics. To help you find the reference of interest we developed an Enzyme Reference Tool that organizes all papers published using our products. With this tool you can filter by quality attribute, enzyme used, type of antibody molecule and technology.

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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.

Find all FAQ here

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.