Genovis / Products / IgG Proteases / GingisKHAN

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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 cystein 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

  • Generation of Fab fragments from human IgG1
  • 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.

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Applications

Applications

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Antibody Characterization

CASSS 2015

"Initial Characterization of a Novel Cysteine Protease for Antibody Subunit Analysis"

FAQ

No, GingisKHAN is a general protease. However, under the recommended reaction conditions, it digests native human IgG1 at one specific site above the hinge.
The enzyme is a cysteine protease that can be inhibited by alkylating agents such as iodoacetamide. There are also several other known inhibitors to this enzyme, like KYT-36. 
The ready-to-use reaction buffer additive has been optimized 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. We recommend preparing a fresh solution of pH neutral cysteine, for use the same day, with a final reaction concentration of 2mM. 
Genovis provides a GingisKHAN Fab kit containing a specific Capture Select CH1 resin for further purification of the Fab. See this page for more information. 

Find all FAQ here

Patent and Disclaimer

GingisKHAN®

The trademark GingisKHAN is the property of Genovis AB.

For research use only. Not intended for any animal or human therapeutic diagnostic use.
©2018 Genovis AB.