FragIT kit

400.004,010.00

FragIT kit easily generates and purifies F(ab’)2 and Fc/2 fragments from IgG

400.00
1,070.00
1,340.00
4,010.00
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Description

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 (other CaptureSelectTM resin needed for rabbit IgG, please inquire). FragIT kit provides digestion and purification 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.

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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