€470.00 – €4,200.00
FabRICATOR Immobilized generates F(ab’)2 and Fc/2 fragments from IgG and Fc-fusion proteins
FabRICATOR Immobilized contains immobilized FabRICATOR (IdeS) enzyme on agarose beads in an easy to use spin column format. FabRICATOR Immobilized 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.
| Application | |
|---|---|
| Product Group | |
| Source/Origin | Cloned from S. pyogenes, Produced in E. coli (His-tagged), 37.7 kDa |
| Product Composition | |
| Digestion Site | |
| Specificity | |
| Substrates | Human IgG1, Human IgG2, Human IgG4, Monkey IgG, Rabbit IgG, Rat IgG, Sheep IgG |
| Reaction Conditions | 30 minute reaction at 37°C according to protocol specifications |
| Digestion Buffers | |
| Buffer Compatibility | |
| Inhibitors | |
| Unit Definition |
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 FabRICATOR Fab2 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 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.
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: ..CPAPELLG / GPSVF..
Human IgG2: ..CPAPPVA / GPSVF..
Human IgG3: ..CPAPELLG / GPSVF..
Human IgG4: ..CPAPEFLG / GPSVF..
