FragIT™ - Immobilized FabRICATOR®

FragIT is FabRICATOR enzyme immobilized on agarose allowing for antibody subunit fragmentation without contaminating the final preparation with FabRICATOR enzyme. The agarose is supplied in spin columns for digestion of small amounts up to hundred milligram of antibody or Fc-fusion protein.

With the FragIT Kit it is possible to go one step further to purify the F(ab')2 from the Fc fragments. The kit consist of two spin column, one with FragIT resin and the other with CaptureSelect™- an agarose matrix with a 13 kDa lama antibody fragment recognizing Fc of multiple species with high affinity.

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FragIT™

FragIT is an extremely rapid and easy way of creating F(ab')2 fragments from IgG.

FragIT is FabRICATOR enzyme immobilized on agarose allowing for antibody subunit fragmentation without contaminating the final preparation with FabRICATOR enzyme.

To view pack sizes and order click here

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FragIT™ Kit

FragIT kit is an extremely rapid and easy way of creating and isolating F(ab')2 and Fc fragments from IgG. The kit consist of two spin columns - one for fragmentation (FragIT column) and one for purification of F(ab')2 and isolation of Fc fragments.

The FragIT column is FabRICATOR enzyme immobilized on agarose and the second column has affinity ligands selective for Fc fragments.

To view pack sizes and order click here

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Products - FragIT™

FragIT - Immobilized FabRCATOR (IdeS) enzyme on agarose in convenient spin columns. The spin columns are available in three different sizes; for digestion of up to 0.5, 10 or 100 mg IgG. The smallest columns are available in three different pack sizes with 2, 5 or 10 columns per pack.

Quotations

Don't hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities.

Products - FragIT™ kit

FragIT Kit contain two different spin columns; FragIT - Immobilized FabRCATOR (IdeS) enzyme on agarose and a CaptureSelect column for purification of F(ab')2 fragments.

FragIT Kit are available in three different sizes; for digestion and purification of up to 0.5, 10 or 100 mg IgG. The smallest columns (for digestion of 0.5 mg IgG) are available in two different pack sizes with 1 or 5 of each column (FragIT and CaptureSelect) per pack.

Note that CaptureSelect columns only are sold together with the FragIT columns as part of these kits and are not available as a stand alone products.

Quotations

Don't hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities.

Detailed Information

FragIT is a resin with FabRICATOR enzyme covalently coupled to agarose beads for subunit fragmentation of IgG to generate pure F(ab’)2 and Fc fragments without contaminating the final preparation with enzyme. IgG is simply incubated with the FragIT resin and fragments are then easily collected by a centrifugation step.

FragIT cleaves IgG at one specific site below the hinge region and thereby eliminating the risk of over digestion if the incubation time is increased.

FragIT digests IgG under physiological reaction conditions thus preserving the immunoreactivity. Digestion is performed at room temperature but if needed (i.e. for mouse IgG) digestion can be preformed at 37°C.

Species

FragIT digests all subclasses of human, monkey, rabbit and sheep IgG. FragIT has limited activity on mouse IgG2a and IgG3 and for digestion of mouse IgG2a and IgG3 the use of FragIT Z is recommended.

Follow this link to view a protease IgG digestion chart with species

Stability

FragIT is supplied in 20% ethanol, shipped on ice and should be stored at +4-8°C upon arrival. Do not freeze the product!

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.

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.

CaptureSelect™ Column

The CaptureSelect columns contains multi species Fc affinity matrix and is an agarose bead with a 13 kDa lama antibody fragment recognizing IgG of multiple species with high affinity. The used ligand is directed towards domains on the Fc part of IgG that enable purification of IgG of, amongst others, human, mouse, bovine, rabbit, rat, goat, horse, and sheep.

CaptureSelect is supplied in 20% ethanol, shipped on ice and should be stored at +4-8°C. Do not freeze the product.

CaptureSelect columns are only sold together with the FragIT columns as part of a kit and are not available as a stand alone products.

Documents

Applications

FabRICATOR in QC applications

FabRICATOR was described in a recent paper by Merck Research Laboratories for domain characterization of therapeutic antibody products. Here you can read more and download the complementary full paper.

mAb Quality in Cell Culture

Due to the extraordinary specificity and robustness of FabRICATOR and IgGZERO they are becoming a vital tool for the ever-increasing demand of analytical characterization of large complex biomolecules.

Find all applications here

Enzyme Reference Tool

FabRICATOR is used frequently for studying important quality attributes of antibody-based biotherapeutics. To help you find the reference of interest we developed an Enzyme Reference Tool that organizes all papers published using our products. With this tool you can filter by quality attribute, enzyme used, type of antibody molecule and technology.

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Latest reference using this product

FAQ and Support

Latest 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

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.

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.

Find all FAQ here

Patent and Disclaimer


FabRICATOR®

Genovis AB hold a license for exclusive worldwide use of biotechnical industrial applications of IdeS covered by any patents derived from International publication WO03051914, including granted US Patent No US7666582.
Other aspects of FabRICATOR technology are encompassed by pending patent applications in the name of Genovis AB.

The trademark FabRICATOR is the property of Genovis AB.


For research use only. Not intended for any animal or human therapeutic diagnostic use.