Below Hinge Digestion of Mutated IgG
FabRICATOR Xtra LALA is an IgG-specific protease that digests antibodies which have been engineered to contain mutated hinges, including the LALA mutation. It digests below the hinge, generating F(ab’)2 and Fc fragments.
FabRICATOR Xtra LALA is active on a variety of antibodies with mutated hinge regions and enables middle-level LC-MS characterization of critical quality attributes of hinge-mutated antibody-based therapeutics.
- Efficient below hinge digestion of hinge-mutated antibodies, including LALA
- Versatile – digests a variety of hinge-mutated antibodies
- Enables middle-level analysis of hinge-mutated antibody therapeutics
Download product folderFabRICATOR® Xtra LALA Lyophilized
Lyophilized enzyme for below hinge digestion of hinge-mutated IgG, including LALAAbout FabRICATOR® Xtra
FabRICATOR Xtra LALA is an enzyme that digests hinge-mutated antibodies, including those containing the LALA mutation.
The enzyme digests below the hinge, generating F(ab’)2 and Fc fragments, which can be reduced into Fd’, LC and scFc fragments. FabRICATOR Xtra LALA digests a number of hinge-mutated antibody formats at two potential sites below the hinge (between residues 235 and 236 or between residues 236 and 237; Table 1).
Table 1. Recommended Reaction Conditions and Digestion Sites for Different Hinge-mutated IgG Formats
| Antibody | Enzyme Amount | Reaction Time | Digestion Site(s) |
| hIgG1-LALA (L234A, L235A) |
1 unit / μg IgG | 1 h | CPAPEAAG / GPSVF (primary), CPAPEAA / GGPSVF (secondary) |
| hIgG4-FALA (F234A, L235A) |
1 unit / μg IgG | 1 h | CPAPEAA / GGPSVF (primary), CPAPEAAG / GPSVF (secondary) |
| hIgG1-LAGA (L235A, G237A) |
2 units / μg IgG | 2 h | CPAPELA / GAPSVF (primary), CPAPELAG / APSVF (secondary) |
| hIgG1-FES (L234F, L235E, P331S) |
3 units / μg IgG | O/N | CPAPEFEG / GPSVF
|
The enzyme is active at pH 7.0-9.0 and at a salt concentration up to 200 mM. No additional cofactors are required for digestion, however, adding a reducing agent (e.g. up to 20 mM DTT) will slightly increase enzymatic activity. The incubation time required for digestion using FabRICATOR Xtra LALA varies depending on the specific hinge mutations present in the antibody (Table 1).
FabRICATOR Xtra LALA is derived from Streptococcus castoreus and expressed in E. coli. The enzyme contains a His-tag and has a molecular weight of 41 kDa.
FabRICATOR® Xtra LALA Lyophilized
Lyophilized enzyme for below hinge digestion of hinge-mutated IgG, including LALA.
FabRICATOR Xtra LALA Lyophilized is available as a lyophilized powder in 2000 unit vials for digestion of up to 2 mg hinge-mutated 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
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Unit Definition
One unit FabRICATOR Xtra LALA Lyophilized digests ≥ 95% of 1 µg human IgG1-LALA (L234A, L235A) when incubated in 10 mM sodium phosphate, 137 mM NaCl, 2.7 mM KCl, pH 7.4 at 37°C for 1 hour.
Content and Storage
FabRICATOR Xtra LALA Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, the enzyme is stable for 1 month at +4-8°C. The product is shipped cold and should be stored at -20°C upon arrival.
Instructions
Product Specifications
Safety Data Sheet
Certificate of Analysis
FAQ and Support
Popular FAQ
Yes, of the formats we have specifically tested, hIgG1-LALAPG (L234A, L235A, P329G), hIgG1-STR (L234S, L235T, G236R) and hIgG1-L234F, L235E, D265A could not be digested to satisfactory levels using FabRICATOR Xtra LALA. Please refer to footnote 4 in the instructions for use to determine whether your antibody is likely to be suitable for FabRICATOR Xtra LALA digestion.
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