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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.
Citations
FAQ and Support
I work with mouse IgG2a containing LC-lambda, can I use another affinity resin for purification of the Fab?
Yes. There is the possibility to exchange the CaptureSelect LC-kappa (mur) columns to columns containing CaptureSelect LC-lambda (mouse). This resin is specific for mouse IgG. Please contact support@genovis.com if this is of interest.
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
How can I stop the FabULOUS reaction?
FabULOUS enzyme reaction is inhibited with iodoacetamide, iodo acetic acid or lowering pH below 3.
Can I digest Fc-fusion proteins using FabULOUS (SpeB)?
Yes, FabULOUS should be able to digest a Fc fusion protein as long as you have not fused the protein too close to the cleavage site. FabULOUS digests human IgG1 just above the hinge (…KTHT / CPPCP…).
Is FabULOUS working on human IgG4?
Yes, but a longer incubation time is needed.
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