FabRICATOR®
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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.
Immobilized enzyme for below hinge digestion of 2 x 0.5 mg IgG in spin columns

Immobilized enzyme for below hinge digestion of 5 x 0.5 mg IgG in spin columns

Immobilized enzyme for below hinge digestion of 10 x 0.5 mg IgG in spin columns

Immobilized enzyme for below hinge digestion of 1-10 mg IgG in spin columns

Immobilized enzyme for below hinge digestion of 10-100 mg IgG in spin columns

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
One spin column contains sufficient FabRICATOR-coupled agarose beads to digest 0.5 mg (Microspin), 10 mg (Midispin), or 100 mg (Maxispin) IgG.
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.
FabRICATOR Immobilized is supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the columns!
Generation and purification of F(ab’)2 and scFc fragments without residual enzyme, enabling rapid preparation of antibody fragments for downstream applications.
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.
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.
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.
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..
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.
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