SmartEnzymes FabULOUS workflow

Above Hinge 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.

The prepared Fab fragments can for example be used in affinity studies, studies of Fab glycosylation and structural studies.
 

  • 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)

FabULOUS Lyophilized

Lyophilized enzyme for above hinge digestion of IgG

FabULOUS Fab Kit

Lyophilized enzyme and affinity resin for above hinge digestion of murine IgG and purification of fragments

About FabULOUS

FabULOUS workflow

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.

FabULOUS Lyophilized

Lyophilized enzyme for above hinge digestion of IgG.
FabULOUS Lyophilized is available as a lyophilized powder in 2000 unit vials for digestion of 2 mg IgG.
 

  • Flexible format for method development
  • Possible to combine enzymes for improved efficiency
  • Amenable to automated workflows
  • Ready to use – just add water!

Available Products

FabULOUS Lyophilized 2000 unitsLyophilized enzyme for above hinge digestion of 2 mg IgGA0-PU1-020607.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit digests ≥ 95% of 1 µg human IgG1 when incubated in PBS with 5 mM DTT or TCEP, pH 7.4 at 37°C for 1 hour.

Content and Storage

FabULOUS Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, FabULOUS is stable for at least 1 month at +4-8°C. The product is shipped cold and should be stored at -20°C upon arrival.

Buffers and Reducing Agents

FabULOUS digests IgG in commonly used buffers with pH ranging from 6.5 to 8.0 and best activity is obtained at 37°C. The enzyme requires reducing agent in the digestion reaction and depending on the reducing conditions, there is a chance that the interchain thiols will be reduced.

Instructions

Product Specifications

Safety Data Sheet

Certificate of Analysis

FabULOUS Fab Kit

Lyophilized enzyme and affinity resin for above hinge digestion of murine IgG and purification of fragments.
FabULOUS Fab Kit contains one vial of FabULOUS enzyme with 2000 units for digestion of 2 mg murine IgG, and 4 CaptureSelect™ LC-Kappa (mur)* microspin columns for affinity purification of fragments.

The affinity matrix recognizes the LC-Kappa domain of murine IgG antibodies, which enables purification of Fab fragments. Each microspin column contains sufficient material to purify 0.5 mg IgG.
 

  • Purification of Fab fragments from murine IgG
  • Fast - pure fragments in under two hours
  • Reliable workflow for preparative applications

Available Products

FabULOUS Fab Kit 2 mgLyophilized enzyme and affinity resin for above hinge digestion of 2 mg murine IgG and purification of fragmentsA1-PFK-0201,017.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit digests ≥ 95% of 1µg human or mouse IgG when incubated in 10 mM Tris, 150 mM NaCl pH 7.4 at 37°C for 1 hour.

Content and Storage

FabULOUS Fab Kit is shipped cold.The FabULOUS Lyophilized vials should be stored at -20°C upon arrival.

The microspin columns containing CaptureSelect™* LC-Kappa (mur) resin should be stored at +4-8°C. Do not freeze the spin columns!

After reconstitution, the FabULOUS enzyme is stable for 1 month at +4-8°C.

Instructions

Product Specifications

Safety Data Sheet

Certificate of Analysis

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

FAQ and Support

Popular FAQ

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.

Yes, but a longer incubation time is needed.

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.

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…).

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.

 

Find all FAQ here

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