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€335.00Add to cart
FragIT Microspin, Digest 5 x 0.5 mg
A0-FR6-025€765.00Add to cart
FragIT Microspin, Digest 10 x 0.5 mg
A0-FR6-050€1,270.00Add to cart
FragIT Maxispin, Digest 10-100 mg
A0-FR6-1000€3,045.00Add 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€380.00Add to cart
FragIT kit, Digest and Purify up to 5 x 0.5 mg
A2-FR2-025€1,020.00Add to cart
FragIT kit Midispin, Digest and Purify 1-10 mg
A2-FR2-100€1,275.00Add to cart
FragIT kit Maxispin, Digest and Purify 10-100 mg
A2-FR2-1000€3,820.00Add 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® 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
We can only guarantee optimal digestion for one time use. However, depending on antibody the columns can be reused but we do not have a proper sanitization protocol. We don’t recommend that you run different antibodies on the same column as we cannot guarantee that there will be no carry-over. If desired to reuse the column store in 10-20% ethanol at +4-8°C.
To use protein A or MabSelect Sure (GE) for removal of Fc as a platform after digestion with FabRICATOR or FragIT is problematic. Therefore we have chosen CaptureSelect in the FragIT Kit for removal of Fc fragments. CaptureSelect is much more specific for Fc. Both protein A and MabSelect Sure (GE) may have affinity for Fab and the degree of affinity is antibody dependent.We have many customer reports on this subject and they usually comes back to us for the CaptureSelect columns for the above reason.
Yes, columns can be reused but make sure you wash the column with buffer. It can be stored in 20% ethanol as well as buffer.
However, we do not have a proper sanitization protocol so we don’t recommend that you run different antibodies on the same column as we cannot guarantee that there will be no carry-over. From an enzymatic activity point of view the column will work for several runs but eventually enzymatic activity will decline.
You can indeed elute the Fc fragments from the CaptureSelect column using a low pH buffer commonly used for example during protein A/G purification of antibodies. We have successfully used 0.1 M glycine buffer pH 2.6-2.8 to release the Fc fragments. Be sure to adjust pH back to normal using a 1 M Tris-HCl buffer or similar directly after isolation of the Fc fragments from the CaptureSelect column.
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For more information about commercial rights, please contact Genovis team at info@genovis.com.
Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals.