Genovis / SmartEnzymes / Antibody Digestion / FabRICATOR / FabRICATOR Immobilized
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FabRICATOR® Immobilized – formerly known as FragIT

Immobilized enzyme for below hinge digestion of IgG in spin columns. FabRICATOR (IdeS) is an IgG-specific cysteine protease that digests antibodies at a single amino acid site below the hinge, generating homogenous F(ab’)2 and Fc fragments.

The FabRICATOR Immobilized spin columns contain FabRICATOR enzyme covalently coupled to agarose beads, for digestion of IgG without contaminating the final preparation with enzyme.

The spin columns are available for digestion of up to 0.5 mg (Microspin), 10 mg (Midispin) or 100 mg (Maxispin) IgG. The smallest columns are available in three different pack sizes, with 2, 5 or 10 columns per pack.
 

  • Easy-to-use spin columns
  • Increased enzyme-to-substrate ratio for improved digestion efficiency
  • No enzyme in the final sample

Available Products

FabRICATOR Immobilized Microspin 2 x 0.5 mgImmobilized enzyme for below hinge digestion of 2 x 0.5 mg IgG in spin columnsA0-FR6-010€456.00Buy / Request a Quote
FabRICATOR Immobilized Microspin 5 x 0.5 mgImmobilized enzyme for below hinge digestion of 5 x 0.5 mg IgG in spin columnsA0-FR6-025€1,028.00Buy / Request a Quote
FabRICATOR Immobilized Microspin 10 x 0.5 mgImmobilized enzyme for below hinge digestion of 10 x 0.5 mg IgG in spin columnsA0-FR6-050€1,695.00Buy / Request a Quote
FabRICATOR Immobilized Midispin 1-10 mgImmobilized enzyme for below hinge digestion of 1-10 mg IgG in spin columnsA0-FR6-100€1,367.00Buy / Request a Quote
FabRICATOR Immobilized Maxispin 10-100 mgImmobilized enzyme for below hinge digestion of 10-100 mg IgG in spin columnsA0-FR6-1000€4,078.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

FabRICATOR Immobilized Citations

Unit Definition

One spin column contains sufficient FabRICATOR-coupled agarose beads to digest 0.5 mg (Microspin), 10 mg (Midispin), or 100 mg (Maxispin) IgG.

Storage and Stability

FabRICATOR Immobilized is supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the columns!

Buffers

FabRICATOR Immobilized can be used with all commonly used buffers with pH ranging from 6.0 to 8.0. Optimization may be required. Note that the FabRICATOR enzyme is irreversibly inactivated below pH 5.0.

FAQ and Support

Popular FAQ

b

Does FabRICATOR introduce modifications?

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.

b

Can I use protein A or MabSelectSure (GE) for removal of Fc-fragments?

Genovis has chosen CaptureSelect in the FabRICATOR Fab2 Kit for removal of Fc fragments. CaptureSelect has a very high specific affinity for the Fc. Both protein A and MabSelect Sure (GE) may have affinity for the Fab which may result in a lower recovery of the generated F(ab’)2 fragments and the degree of affinity is antibody dependent. We have many customer reports on this subject, and they usually come back to us for the CaptureSelect columns for the above reason.

b

Can I re-use FabRICATOR Immobilized columns?

We can only guarantee optimal digestion for one-time use. We do not have a cleaning or regeneration protocol to provide. However, depending on the antibody and the following application the column can be reused. We do not recommend using with different antibodies due to the risk for contamination of carry-over from previous sample. If column reuse is desired store the column in 10-20% ethanol at +4-8°C.

b

Where does FabRICATOR cleave the IgG molecule?

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:  ..CPAPELLG / GPSVF..
Human IgG2:  ..CPAPPVA / GPSVF..
Human IgG3:  ..CPAPELLG / GPSVF..
Human IgG4:  ..CPAPEFLG / GPSVF..

 

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