GingisKHAN® (Kgp)
GingisKHAN (Kgp) is a cysteine protease that digests human IgG1 at a specific site above the hinge. The enzyme requires only mild reduced condition for activity and the reaction is performed in the presence of 2 mM cysteine at neutral pH.
GingisKHAN is available in two product formats:
- GingisKHAN Enzyme - lyophilized enzyme and reducing agent
- GingisKHAN Fab kit - lyophilized enzyme and reducing agent together with affinity purification resin for easy generation and purification of Fab fragments from human IgG1.
Product Information
GingisKHAN is a cysteine protease that digests human IgG1 at a specific site above the hinge (KSCDK / THTCPPC), generating intact Fab and Fc fragments. A second digestion site on the Fc may appear if the N-glycans are removed. GingisKHAN requires mild reducing conditions to be active, and optimal activity is obtained at 37°C and pH 8. The reducing agent (2 mM cysteine) is provided together with the enzyme.
GingisKHAN is purified from Porphyromonas gingivalis.
Key Characteristics
- Human IgG1
- 60 min reaction
- 2 mM cysteine (included)
- KSCDK / THTCPPCP (above the hinge)
Applications
- Subunit LC-MS analysis
- Characterize heavy and light chain pairing of bispecific antibodies
- Study monovalent binding of Fab fragments to the antigen
- Epitope mapping using HDX-MS
- Cystallization of Fab fragments
Product Formats
GingisKHAN® Enzyme
GingisKHAN enzyme supplied as lyophilized powder together with 5 vials of lyophilized GingisKHAN reducing agent for convenient and reliable performance. The enzyme is available in 2000 unit vials for digestion of up to 2 mg human IgG1.
GingisKHAN® Fab kit
GingisKHAN Fab kit is used for rapid and easy preparation and purification of of Fab fragments from human IgG1 antibodies. The kit consists of two parts; lyophilized GingisKHAN enzyme for digestion of human IgG1 and affinity spin columns for purification of intact Fab fragments.
Product Documents
FabALACTICA Brochures, Posters and Applications
Brochures and Posters
Download Genovis brochures and posters with the latest details on the SmartEnzymes
Download GingisKHAN Poster and External References
Poster from AT Europe 2014
"Initial Characterization of a Novel Cysteine Protease for Antibody Subunit Analysis"
FAQ and Support
Popular FAQ
Yes, GingisKHAN should be able to digest a Fc fusion protein if you have not fused the protein too close to the cleavage site and the Fc-part is from a human IgG1. GingisKHAN digests human IgG1 just above the hinge (…KSCDK / THTCPPCP…). If there are exposed Lysines in the fusion part, digestion might occur there as well.
No. GingisKHAN is a protease that digests at exposed Lysines with no secondary structure restrictions. At the recommended reaction conditions, GingisKHAN digestion above the hinge has only been confirmed on human IgG1 with the exposed lysine above the hinge, (…KSCDK / THTCPPCP…). No digestion has been confirmed on other subclasses of human IgG except for several digestion sites in human IgG3.
Yes. GingisKHAN is provided lyophilized. After reconstitution, the enzyme can be used for up to 4 different antibodies. The digestion is performed in solution and you can prepare Fab fragments from 4 x 0.5 mg human IgG1. The purification step is done with Capture Select columns and 4 columns are provided in the kit.
We recommend using the 10x reducing agent provided with GingisKHAN which has an optimized pH to provide full enzymatic activity without affecting the disulphide bridges between the light chain and Fd region of the heavy chain, keeping the Fab intact. If other reducing agent is used there is a risk for non-optimal reaction conditions. It is important to use freshly prepared Reducing Agent (max storage 6h) for guaranteed activity of GinigsKHAN.
No, GingisKHAN is a general protease that with the recommended reaction conditions, 1 hour with light reducing conditions, digests human IgG1 at one specific site (…KSCDK/THTCPPCP…) above the hinge, generating Fab and Fc.
