FabALACTICA™ – Above Hinge Digestion of Human IgG1

FabALACTICA (IgdE) is a cysteine protease that digests human IgG1 at a specific site above the hinge, generating intact and homogenous Fab and Fc fragments.

SmartEnzymes™

FabALACTICA facilitates the characterization of bi- and multispecific antibodies and analyses of intact and paired Fc glycosylation. It also simplifies the study of monovalent binding, higher order structures, disulfide scrambling and antibodies with mutated hinge regions.

Why FabALACTICA™?

  • Specific – one digestion site above the hinge of human IgG1
  • Generates homogeneous Fab and Fc fragments with preserved immunoreactivity
  • Middle-level characterization of human IgG1
  • Available immobilized in ready-to-use spin columns

  • Target

    Human IgG1

  • Time

    O/N (16-18 hr) reaction

  • Box

    No need for reducing agents or co-factors

  • Digestion Site

    KSCDKT / HTCPPCP (above the hinge)

Product Formats

About FabALACTICA™

FabALACTICA is a cysteine protease that digests human IgG1 at one specific site above the hinge (KSCDKT / HTCPPC), generating intact Fab and Fc fragments.

The name of the enzyme is derived from the pathogen Streptococcus agalactiae, where it was first discovered (Spoerry et al., 2016). The enzyme is active at pH from 6-8 and does not require reducing conditions or co-factors for activity.

FabALACTICA is recombinantly expressed in E. coli, contains a His-tag, and the molecular weight is 70 019 Da.

Spoerry, C. et al., 2016. Novel IgG-Degrading Enzymes of the IgdE Protease Family Link Substrate Specificity to Host Tropism of Streptococcus Species. PloS one, 11(10), p.e0164809.

Performance Activity

Citations

Resources

FAQ and Support

Yes, for human IgG1 molecules both enzymes can be used in combination to generate a specific hinge peptide. The incubation time is significantly longer for FabALACTICA and it is recommended to first run the FabALACTICA digest and then add FabRICATOR for the last 30 min.

We recommend using FabALACTICA Immobilized for digesting human IgG1 and generating a final preparation free of any residual enzyme. You can also use FabALACTICA Fab kit for digestion of human IgG1 and purification of the generated Fab fragments.

No, FabALACTICA has shown very poor activity on Fc-fusion proteins.

FabALACTICA (IgdE) and GingisKHAN (Kgp) are both cysteine proteases that digest IgG1 above the hinge to generate intact Fab and Fc fragments. FabALACTICA is specific for human IgG1 and digests at a specific site (…KSCDKT / HTCPPCP…) with overnight incubation without the need for reducing conditions. GingisKHAN is a general protease that with the recommended reaction conditions, 1 hour with light reducing conditions, digests human IgG1 at a specific site (…KSCDK/THTCPPCP…).

We can only guarantee optimal digestion for one-time use. We do not have a cleaning or regeneration protocol to provide. However, depending on the antibody and the following application the column can be reused. We do not recommend using with different antibodies due to the risk for contamination of carry-over from previous sample. If column reuse is desired store the column in 10-20% ethanol at +4-8°C.

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