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Endo-beta-N-acetylglucosaminidase EndoS2 mutant

Endo-beta-N-acetylglucosaminidase EndoS2、Endoglycosidase S2、EndoS2

价格 ￥4,860.00 （ 供应商现货 ： 3-5个工作日）
货号 UA070139
规格
数量
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产品介绍

产品介绍 评论(0)

产品规格

  • 物种

    Streptococcus pyogenes

  • 分子别名

    Endo-beta-N-acetylglucosaminidase EndoS2、Endoglycosidase S2、EndoS2

  • 表达宿主

    E.coli

  • 分子量

    93 Da (Reducing)

  • 纯度

    ＞95% by SDS-PAGE

  • 活性

    2.32mg/mL

  • 标记

    Unconjugated

  • 标签

    His Tag

  • 缓冲体系

    20 mM Tris-HCl、50 mM NaCl、pH 7.5 @ 25°C

  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    [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.
    [2] Li T, et al. "Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. " J Biol Chem. 2016 Aug 5;291(32):16508-18.

背景介绍

  • 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.

产品组分

  • 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)

操作步骤

  • 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

注意事项

  • Please avoid repeated freeze-thaw cycles.

酶活定义
  • 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.

  • 生物活性

    • 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

  • 电泳

    • 2μg (R: reducing condition, N: non-reducing condition).

  • 反相高效液相色谱(RP-HPLC)

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