Genovis / SmartEnzymes / Antibody Digestion / FabRICATOR / FabRICATOR Fab2 Kit
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FabRICATOR® Fab2 Kit – formerly known as FragIT kit

Immobilized enzyme and affinity resin for below hinge digestion of IgG and purification of fragments. 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.

FabRICATOR Fab2 Kit consists of spin columns of FabRICATOR Immobilized for antibody digestion, and spin columns of CaptureSelect™* Fc resin for affinity purification of F(ab’)2 and Fc fragments. One spin column contains sufficient FabRICATOR-coupled agarose beads to process 0.5 mg (Microspin), 10 mg (Midispin), or 100 mg (Maxispin) IgG.

FabRICATOR Fab2 Kit is available for digestion and purification of 0.5 mg - 100 mg antibody or Fc-fusion protein. It works on all subclasses of human, and some classes of monkey, rabbit and sheep IgG, but has limited activity on mouse IgG2a and IgG3 - for digestion and purification of these antibody species, we recommend using FabRICATOR Z Fab2 Kit (formerly FragIT Z kit).
 

  • Purification of homogeneous F(ab’)2 fragments from several IgG species and subclasses
  • Fast - pure fragments within 60-90 minutes
  • Reliable workflow for preparative applications

Available Products

FabRICATOR Fab2 Kit Microspin 0.5 mgImmobilized enzyme and affinity resin for below hinge digestion of 0.5 mg IgG and purification of fragmentsA2-FR2-005€529.00Buy / Request a Quote
FabRICATOR Fab2 Kit Microspin 5 x 0.5 mgImmobilized enzyme and affinity resin for below hinge digestion of 5 x 0.5 mg IgG and purification of fragmentsA2-FR2-025€1,408.00Buy / Request a Quote
FabRICATOR Fab2 Kit Microspin 10 x 0.5 mgImmobilized enzyme and affinity resin for below hinge digestion of 10 x 0.5 mg IgG and purification of fragmentsA2-FR2-050€2,683.00Buy / Request a Quote
FabRICATOR Fab2 Kit Midispin 10 mgImmobilized enzyme and affinity resin for below hinge digestion of 10 mg IgG and purification of fragmentsA2-FR2-100€1,769.00Buy / Request a Quote
FabRICATOR Fab2 Kit Maxispin 100 mgImmobilized enzyme and affinity resin for below hinge digestion of 100 mg IgG and purification of fragmentsA2-FR2-1000€5,272.00Buy / Request a Quote

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

FabRICATOR Fab2 Kit 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

The FabRICATOR Immobilized and CaptureSelect™* spin columns are 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.

* Thermo Scientific™ CaptureSelect™ resin from Thermo Fisher Scientific Inc. and its subsidiaries. Thermo Fisher and CaptureSelect are trademarks of Thermo Fisher Scientific Inc. and its subsidiaries.

FAQ and Support

Popular FAQ

b

Can I release Fc fragments from the CaptureSelect column after isolating the F(ab’)2 fragments?

It is possible to elute the Fc fragments from the CaptureSelect column using a low pH buffer, for example a 0.1M Glycine buffer pH=3.0. The pH must immediately after elution be adjusted to neutral pH by adding 0.1x elution volume of 1M Tris pH=8.0.

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

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 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

Can I use FabRICATOR Fab2 Kit for generation and purification of fragments from Rabbit IgG?

FabRICATOR Fab2 Kit digests rabbit IgG but to capture the Fc fragments in the affinity step you need another resin than the CaptureSelect™ Fc normally included in the kit. Please contact the Genovis team if you are interested in processing rabbit IgG and we will provide columns with resin compatible with rabbit Fc in the FabRICATOR Fab2 Kit.

 

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