Product Overview
The Hzymes Endo S2 Pro is a recombinant glycosidase derived from the glycosidase of Streptococcus pyogenes through sequence redesign and mutation modification. It combines the endoglycosidase functions of EndoS and EndoH, is expressed in Escherichia coli, and has no purification tags.
It can specifically cleave the β (1-4) glycosidic bonds between the first and second N-acetylglucosamine residues near the Asp on the N-glycan chain of IgG, including complex-type, heterozygous and high-mannose types, etc. And it exhibits high glycosylation activity and can utilize oxazoline sugars as glycosyl donors for antibody glycan conjugation.
The purity of this product is approximately 95%, with high activity, good stability, free of exoglycosidase contamination, and no protein hydrolysis activity. It demonstrates superior flexibility and efficiency in antibody glycosylation modification, glycan chain analysis, and glycoengineering research.
Key Features
Proprietary Engineered Enzyme
Derived from Streptococcus pyogenes glycosidase through sequence redesign and mutation optimization, with independent intellectual property and no known patent restrictions.
Broad Substrate Compatibility
Combines functional characteristics of EndoS and EndoH, enabling cleavage of β (1-4) glycosidic bonds in IgG N-glycans, including complex-type, hybrid, and high-mannose glycans under defined conditions.
High Purity and Tag-Free Design
Expressed in E. coli without purification tags. Typical purity is approximately 95%, as determined by standard analytical methods.
Good Enzymatic Activity and Stability
Demonstrates consistent glycosidase activity under recommended conditions, with no detectable exoglycosidase contamination or protein hydrolysis activity.
Defined Specific Activity
Enzyme specific activity is >50 kU/mg under standard assay conditions.
Applications
Antibody Glycan Engineering
Used for enzymatic remodeling of IgG N-glycans, supporting antibody structure-function studies and glycoengineering workflows.
Antibody Conjugation and Labeling
Applicable in bioconjugation strategies involving glycan-based modification of antibodies for downstream functional studies or assay development.
Protein Modification and Cross-Linking
Suitable for controlled modification of glycoproteins, including labeling, immobilization, and cross-linking applications in biochemical research.
Glycan Analysis and Characterization
Used in structural analysis of antibody glycosylation profiles, supporting comparative studies of different glycoforms.
Performance Data
One unit is defined as the amount of enzyme required to remove 95% of the glycan chains from 5 μg of IgG in a 10 μl reaction system at 37°C within 1 hour.
Comparative Result
Activity Identification
Lane 1-4: Enzymatic cleavage activity of wt (wild-type enzyme) at enzyme: IgG mass ratios of 1:100, 1:250, 1:500, and 1:1000 respectively.
Lane 5-8: Enzymatic cleavage activity of Endo S2 Pro at enzyme: IgG mass ratios of 1:100, 1:250, 1:500, and 1:1000 respectively.
Lane 9: Fc fragment of IgG digested with IdeZ enzyme.
FAQ
What is Endo S2 Pro and what is its function in ADC development?
Endo S2 Pro is a recombinant engineered endoglycosidase derived from Streptococcus pyogenes glycosidase through sequence redesign and mutation optimization. It is expressed in E. coli without any purification tag and combines functional characteristics of EndoS and EndoH.
In ADC (antibody-drug conjugate) development, Endo S2 Pro is mainly used for:
• Glycan remodeling and deglycosylation: It cleaves β (1-4) glycosidic bonds between N-acetylglucosamine residues in IgG Fc N-glycans, including complex-type, hybrid-type, and high-mannose glycans under defined conditions.
• Glycan-based conjugation: Enables glycan-directed modification of antibodies for site-specific conjugation and bioconjugation workflows.
The enzyme is designed to support flexible glycoengineering strategies, including glycan chain analysis and antibody modification applications.
What are the key advantages of Endo S2 Pro?
Endo S2 Pro offers several features relevant to glycoengineering applications:
• Engineered design with IP protection: Developed through sequence redesign and mutation optimization with independent intellectual property.
• Broad substrate compatibility: Retains activity toward multiple IgG glycoforms, including complex, hybrid, and high-mannose types under suitable conditions.
• Tag-free recombinant expression: Produced in E. coli without purification tags, supporting downstream compatibility in protein workflows.
• Good enzymatic performance and stability: Shows consistent activity under recommended conditions, with low contamination of exoglycosidase activity and no detectable protein hydrolysis activity.
• Defined specific activity: Enzyme specific activity is >50 kU/mg under standard assay conditions.
What glycoengineering applications is Endo S2 Pro used for?
Endo S2 Pro is used in a range of antibody and glycoprotein engineering workflows, including:
• Antibody glycan remodeling: Modification of IgG Fc glycans for structure-function studies and glycoengineering research.
• Antibody conjugation and labeling: Supports glycan-directed site-specific conjugation strategies for antibodies and related biomolecules.
• Protein modification and immobilization: Used in controlled protein engineering workflows such as cross-linking, labeling, and immobilization.
• Glycan chain analysis: Applied in analytical workflows for characterization of antibody glycosylation profiles.
What is the difference between Endo S2 Pro and wild-type Endo S2?
Endo S2 Pro is an engineered variant developed through sequence redesign and mutation modification based on wild-type Endo S2.
Compared with the wild-type enzyme, Endo S2 Pro is designed to provide improved compatibility in glycoengineering workflows while addressing known intellectual property constraints associated with the native enzyme. It is intended for research and development applications in antibody engineering and related fields.
