FragIT

350.003,200.00

FragIT generates F(ab’)2 and Fc/2 fragments from IgG and Fc-fusion proteins

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3,200.00
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Description

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.

Detailed Information

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FAQs

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.

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.

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.

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

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