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IgASAP™ – Digestion of IgA
IgASAP is a family of IgA-digesting enzymes. IgASAP Sub1 is an IgA1-specific enzyme that digests human IgA1 at one specific site above the hinge, and IgASAP Sub1+2 specifically digests IgA1 and IgA2m1 above the hinge. Both enzymes generate intact and homogenous Fab and Fc fragments.
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The IgASAP enzymes enable generation of intact monovalent Fab fragments as well as middle-level analysis of IgA, which facilitates IgA characterization during, for example, the development of vaccines, therapeutics and diagnostics.
Why IgASAP™?
- Unique enzymatic tools that enable middle-level characterization and quality control of IgA
- Generates homogeneous Fab and Fc fragments with preserved immunoreactivity
- Highly specific – one digestion site above the hinge of human IgA
Sub1: Human IgA1 Sub1+2: Human IgA1 and IgA2m1
Sub1: 1 hour reaction Sub1+2: O/N (16-18 h) reaction
No need for reducing agents or co-factors
Above the hinge (see digestion sites below)
Product Formats
IgASAP™ Sub1+2 Lyophilized
Lyophilized enzyme for above hinge digestion of human IgA1 and IgA2m1
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About IgASAP™
The IgASAP family of IgA-digesting enzymes specifically digest serum and secretory type human IgA to generate Fab and Fc fragments. IgASAP Sub1 specifically digests human IgA1 while IgASAP Sub1+2 digests both human IgA1 and IgA2m1.
As an essential component of the immune system, IgA plays a role in many different health conditions from autoimmune disease, allergic reactions, gastrointestinal disorders and others. The ability to selectively and specifically digest IgA under native conditions and in complex biological samples can be of great value in clinical medicine and research. The IgASAP enzymes are valuable tools to reduce the sample complexity of therapeutic IgA candidates and significantly simplify analytical characterization of IgA.
One significant structural difference between human IgA1 and IgA2 is the hinge region. IgA1 has a longer O-glycan-rich and more flexible hinge region compared to IgA2. The digestion site of IgASAP Sub1 is located in this extended region, rendering the enzyme IgA1-specific.
The IgA2 subclass exists in three allelic forms, A2m1, A2m2 and A2m3. IgASAP Sub1+2 digests IgA1 and IgA2m1 between proline (P) and valine (V) just N-terminal to the hinge region, but since the proline residue in A2m2 and A2m3 is substituted with an arginine (R) residue, these two allotypes of IgA2 are resistant to digestion.
Applications
Site-specific digestion of human IgA1, generating homogeneous Fab and Fc fragments for high-resolution middle-level LC-MS characterization.
Site-specific digestion of human IgA2m1, generating homogeneous Fab and Fc fragments for high-resolution middle-level LC-MS characterization.
Subclass selective digestion of IgA in complex samples like serum, enabling targeted analysis of IgA1 and IgA2m1 without off-target cleavage.
Performance Activity
Highly specific digestion of IgA, with no activity on IgG or IgM, enabling targeted middle-level characterization of IgA1 and IgA2 subclasses.
Citations
FAQ and Support
Is there an enzyme for digestion of IgA2-only?
There is no known enzyme that targets IgA2-only since the amino acids in the hinge regions of IgA2 is present also in IgA1. However, IgASAP Sub1 will digest IgA1-only.
Does IgASAP Sub1 digest human IgA1-only?
IgASAP Sub1 may digest IgA1 from other primate species with similar hinge sequences, but this has not been tested. IgASAP Sub1 does not digest IgA2.
Can the IgASAP Sub1 digestion be performed at lower temperatures?
The optimal activity is at 37°C, but the digestion can also be performed at room temperature using a prolonged incubation time.
What enzyme should be selected for the digestion of IgA1?
IgASAP Sub1 specifically targets IgA1 and exhibits a faster reaction time (1 hour) compared to IgASAP Sub1+2 (16-18 hours). The cleavage site of IgASAP Sub1+2 is located six amino acids above that of IgASAP Sub1, resulting in the absence of potentially O-glycosylated residues on the Fab fragments. This characteristic can be advantageous in certain cases.
Which buffers are compatible with IgASAP Sub1?
The enzyme is active at pH 6.5 to 8.0, tolerates up to 300 mM NaCl, and does not require reducing conditions or co-factors for activity. PBS and TBS buffers are both compatible. Optimization may be required if buffers other than the recommended are used.
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