FragIT™
The FragIT spin columns contain FabRICATOR (IdeS) enzyme covalently coupled to agarose beads, for digestion of IgG without contaminating the final preparation with enzyme.
FabRICATOR (IdeS) is a cysteine protease that digests antibodies at a specific site below the hinge, generating a homogenous pool of F(ab')2 and Fc/2 fragments after 30 min incubation.
Detailed Information
FabRICATOR digests IgG at a specific site below the hinge region, generating a homogenous pool of F(ab')2 and Fc/2 fragments. The IgG is digested within 30 minutes, and there is no risk of overdigestion if the incubation time is prolonged. Since FabRICATOR digests IgG under physiological reaction conditions, the immunoreactivity is preserved.
FragIT is a resin with FabRICATOR enzyme covalently coupled to agarose beads for generation of F(ab’)2 and Fc/2 fragments without contaminating the final preparation with enzyme. The workflow is simple - IgG is incubated with the FragIT resin and fragments are easily collected by a centrifugation step. 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.
FragIT digests all subclasses of human, and some classes of monkey, rabbit and sheep IgG. It has limited activity on mouse IgG2a and IgG3 - for digestion of these antibody species, we recommend using FragIT Z .
Storage and Stability
The FragIT spin columns are supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the products!
Buffers
FragIT 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.
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.
FragIT - Store
FragIT contains immobilized FabRICATOR (IdeS) enzyme on agarose beads in an easy to use spin column format. FragIT digests antibodies or Fc-fusion proteins at one single amino acid position below the hinge region generating Fc and F(ab’)2 fragments. FragIT is provided in spin columns for digestion of 0.5 – 100 mg of antibody or Fc-fusion protein without residual enzyme in the final preparation.
FragIT Microspin, Digest 2 x 0.5 mg
A0-FR6-010€390.00Add to cart
FragIT Microspin, Digest 5 x 0.5 mg
A0-FR6-025€880.00Add to cart
FragIT Microspin, Digest 10 x 0.5 mg
A0-FR6-050€1,450.00Add to cart
FragIT Midispin, Digest 1-10 mg
A0-FR6-100€1,170.00Add to cart
FragIT Maxispin, Digest 10-100 mg
A0-FR6-1000€3,490.00Add to cart
FragIT Kit - Store
FragIT kit contains immobilized FabRICATOR (IdeS) enzyme on agarose beads in spin columns and CaptureSelectTM for affinity capture of Fc fragments. FragIT digests antibodies or Fc-fusion proteins at one single amino acid position below the hinge region generating Fc and F(ab’)2 fragments. CaptureSelectTM spin columns capture Fc fragments for purification of F(ab’)2 and Fc fragments. FragIT kit provides digestion and purification of 0.5 – 100 mg of antibody or Fc-fusion protein without residual enzyme in the final preparation.
FragIT kit, Digest and Purify up to 0.5 mg
A2-FR2-005€440.00Add to cart
FragIT kit, Digest and Purify up to 5 x 0.5 mg
A2-FR2-025€1,170.00Add to cart
FragIT kit, Digest and Purify up to 10 x 0.5 mg
A2-FR2-050€2,230.00Add to cart
FragIT kit Midispin, Digest and Purify 1-10 mg
A2-FR2-100€1,470.00Add to cart
FragIT kit Maxispin, Digest and Purify 10-100 mg
A2-FR2-1000€4,380.00Add to cart
How to Order
For details on ordering please visit How to order. Click add to cart to buy online.
Quotations
Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities.
Documents
Instructions
Product Specifications
Safety Data Sheet
Certificate of Analysis
FAQ and Support
Popular FAQ
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.
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 FragIT 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: HTCPPCPAPELLG / GPSVF
Human IgG2: VECPPCPAPP_VA / GPSVF
Human IgG3: PPCPRCPAPELLG / GPSVF
Human IgG4: PHAHHAQAPEFLG / GPSVF
