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GingisKHAN® Fab kit

Use GingisKHAN Fab kit to generate purified and homogenous Fab fragments. The kit involves digestion of human IgG1 with GingisKHAN and purification of the Fab fragments on CH1 specific affinity spin columns.

Products

GingisKHAN Fab kit contains one vial of GingisKHAN enzyme with 2000 units for digestion of up to 2 mg human IgG together with 5 vials of lyophilized GingisKHAN reducing agent and 4 CH1 micro spin columns where each column includes sufficient material to purify up to 0.5 mg IgG.

Quotation & ordering

Don't hesitate to contact us if you would like to receive a quotation.
Please find information for placing orders at How to Order.

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Detailed Information

GingisKHAN Fab kit is for rapid and easy preparation and purification of Fab fragments from human IgG1 antibodies. The kit contains of GingisKHAN enzyme for digestion of human IgG1 and IgG-CH1 specific affinity spin columns for purification of the Fab fragments.

GingisKHAN digests human IgG1 at one single site in the upper hinge (…KSCDK / THTCPPCP…) yielding intact Fab and Fc fragments.  A second cleavage site on the Fc may appear if the N-glycans are removed. Intact Fab and Fc fragments are obtained with GingisKHAN digestion of human IgG1 since mild reducing conditions (i.e. 2 mM cysteine) is sufficient for enzyme activity. Optimal activity is obtained at 37°C and pH 8. Reducing agent is supplied with the enzyme.

The CaptureSelect™ IgG-CH1* affinity matrix recognizes the CH1 domain of human IgG antibodies, which enables purification of Fab fragments, independent of the light-chain isotype and source material. Due to its unique selectivity for the CH1 domain, no co-purification of free light-chain contaminants will occur.

Storage

GingisKHAN Fab kit is shipped on ice.

GingisKHAN 2000u and GingisKHAN Reducing Agent should be stored at -20°C upon arrival. CaptureSelect IgG-CH1 resin* in MicroSpin columns should be stored in +4-8°C.

After reconstitution of GingisKHAN enzyme, it is stable for 2 months at +4-8°C. Reconstituted GingisKHAN Reducing Agent should be used the same day as prepared, it cannot be stored.

Unit definition

One unit of GingisKHAN enzyme digests ≥ 95% of 1μg human IgG1 when incubated in 0.1 M Tris, pH 8.0 at 37°C for 1h.

Applications

Purified and homogenous Fab fragments can be used in many applications, including:

  • Study monovalent binding of Fab fragments to the antigen
  • Characterize heavy and light chain pairing of bispecific antibodies
  • Epitope mapping using HDX-MS
  • Cystallization of Fab fragments

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Digestion and purification of Fab fragments using GingisKHAN FabKit. (A) Intact monoclonal IgG1 antibody. (B) Analysis of antibody fragments after GingisKHAN digestion. (C) The digestion mixture was added to the CH1 specific CaptureSelect column and the flow-through Fc fragment is shown. (D) Elution of the purified Fab fragments. Digestion and purification were analyzed on RP-UHPLC (Agilent 1260) using Supelco C4 bioshell A4.

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SDS-PAGE analysis of purified Fab fragments from monoclonal hIgG1 Herceptin using GingisKHAN Fab kit. Lane 1 and 6; MW marker, Lane 2; intact human IgG1, lane 3: Fab and Fc fragments after GingisKHAN digestion, lane 4: flow-through Fc fragments and GingisKHAN, lane 5: eluted Fab fragments.

Documents

Enzyme Reference Tool

All published papers on how Genovis enzymes help simplify antibody analytics. With this tool you can filter by quality attribute, enzyme used, type of antibody molecule and technology.

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FAQ and Support

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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.
*Made with Thermo Scientific CaptureSelect resin from Thermo Fisher Scientific Inc. and its subsidiaries. Thermo Scientific and CaptureSelect are trademarks of Thermo Fisher Scientific Inc. and its subsidiaries.