The results of 5μg substrate digestion separated under diferent quantity of UA070139, the reaction was incubated for 1 h at 37°C.
M marker
Lan1 human IgG 5μg + 232ng UA070139
Lan2 human IgG 5μg + 58ng UA070139
Lan3 human IgG 5μg + 29ng UA070139
Lan4 human IgG 5μg + 12ng UA070139
Lan5 human IgG 5μg + 6ng UA070139
Lan6 human IgG 5μg + 3ng UA070139
Lan7 human IgG 5μg
Endo-beta-N-acetylglucosaminidase EndoS2 mutant
Endo-beta-N-acetylglucosaminidase EndoS2 mutant
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Product Details
Product Details
Product Specification
| Species | Streptococcus pyogenes |
| Synonyms | Endo-beta-N-acetylglucosaminidase EndoS2、Endoglycosidase S2、EndoS2 |
| Expression System | E.coli |
| Molecular Weight | 93 Da (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Storage Buffer | 20 mM Tris-HCl、50 mM NaCl、pH 7.5 @ 25°C |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | [1] Sjögren Jonathan, et al. "EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans." Glycobiology 10:1053-1063. |
Background
EndoS2 is an endo-β-N-acetylglucosaminidase derived from Streptococcus pyogenes. It hydrolyzes the chitobiose core of N-linked glycans, cleaving between the two innermost GlcNAc residues. A key advantage of EndoS2 is its remarkably broad substrate specificity; unlike the original EndoS, it efficiently processes high-mannose, hybrid, and complex-type glycans. This makes it a universal tool for glycan remodeling across various species, including human, mouse, rat, and other mammalian IgGs.
To enhance its synthetic utility, a representative glycosynthase mutant has been engineered. This mutation diminishes the enzyme's hydrolytic activity while retaining its ability to utilize an activated sugar oxazoline donor. This modification shifts the reaction equilibrium overwhelmingly toward synthesis, enabling the production of homogeneous glycoproteins with high yield.
This mutant is particularly valuable in therapeutic antibody development. It facilitates the creation of defined glycoforms to enhance effector functions like ADCC. Consequently, the EndoS2 mutant system provides a robust platform for producing next-generation biotherapeutics with improved efficacy.
Components
Storage Solution : 2.32mg/mL EndoS2 mutant、20 mM Tris-HCl, 50 mM NaCl. (pH 7.5)
10*Reaction Buffer： 500 mM sodium acetate (pH 6.0)
Protocol
1. Combine 100 µg of native IgG, 1 µl of 10*Reaction Buffer and ddH20 (if necessary) to make a 10 µl total reaction volume.
2. Add 1 µl EndoS2.
3. Incubate reaction at 37°C for 1 hour
4. Add 1 µl EndoS2 mutant(The molar ratio of donor to acceptor was 20:1)
5. Incubate reaction at 37°C for 1 hour
Guidelines
Please avoid repeated freeze-thaw cycles.
Unit Definition
One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 5 μg of native mouse monoclonal IgG in 1 hour at 37°C in a total reaction volume of 10 µl.
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Bioactivity
SDS-PAGE
2μg (R: reducing condition, N: non-reducing condition).
RP-HPLC
