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FabULOUS™ – Hinge-region Digestion of IgG
FabULOUS (SpeB) is a cysteine protease that digests in the hinge region of IgG from several different species and subclasses, generating Fab and Fc fragments.
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After FabULOUS digestion, the prepared Fab fragments can for example be used in affinity studies, studies of Fab glycosylation and structural studies.
Why FabULOUS™?
- Active on IgG from multiple species and subclasses – digests in the upper-hinge region
- Fast – generates Fab and Fc fragments within one hour
- Efficient for digestion of mouse IgG1
Human IgG1-4, IgG from mouse, rat, goat, sheep and rabbit
60 min reaction
Requires reducing conditions
Human IgG1: KTHT / CPPCPAP (above the hinge)
Product Formats
FabULOUS™ Fab Kit
Lyophilized enzyme and affinity resin for hinge-region digestion of murine IgG and purification of fragments
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About FabULOUS™
FabULOUS (SpeB) is an enzyme for digestion of IgG in the hinge region. Human, mouse, rat, goat and sheep IgG are digested by FabULOUS, yielding Fab and Fc fragments. The primary digestion site on human IgG1 is …KTHT / CPPCPAPE…
FabULOUS is derived from Streptococcus pyogenes and is expressed in E. coli. The molecular weight of FabULOUS is 29 kDa and the enzyme contains a His-tag.
Performance Activity
Preparation of pure, homogeneous mouse IgG Fab fragments with no residual enzyme by combining FabULOUS digestion and affinity purification.
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FAQ and Support
Is FabULOUS working on human IgG4?
Yes, but a longer incubation time is needed.
Will the reducing conditions required for digestion impact the interchain disulfide bond integrity?
Depending on what reducing agent and what concentration you use you can obtain intact Fab and not reduce the interchain disulfide bond. The light chain (LC) and heavy chain (HC) will not dissociate under native conditions due to hydrogen/hydrophobic forces. Before you analyze your samples with SDS-PAGE you must remove the cysteine otherwise the Fab will be reduced to Fd and LC during sample preparation for the SDS-PAGE. This can be done by a simple buffer exchange.
Can I purify Fab fragments from human IgG with FabULOUS Fab kit?
No, FabULOUS Fab kit contains CaptureSelect LC-kappa (mur) spin columns with selective affinity for murine LC-kappa. If you need to generate pure Fab from human IgG1 you can use either FabALACTICA Fab Kit or GingisKHAN Fab Kit. If you have a different subclass of human IgG, please contact support@genovis.com for guidance.
How can I stop the FabULOUS reaction?
FabULOUS enzyme reaction is inhibited with iodoacetamide, iodo acetic acid or lowering pH below 3.
Does FabULOUS digest all human IgG subclasses above the hinge?
FabULOUS is a cysteine protease that digests IgG in the hinge region. A primary digestion site for human IgG1 is above the hinge (…KTHT / CPPCP…). The enzyme is also active on human IgG2 and IgG3, digestion site not determined but Fab and Fc is generated. FabULOUS digests human IgG4 primarily at a site below the hinge using significantly longer incubation, generating Fab’ due to the reducing conditions. The possibility of other, secondary digestions sites is antibody dependent
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