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Hydrolysis of Flexible Linkers by GlySERIAS

GlySERIAS is a unique enzyme that digests flexible glycine-rich fusion protein linkers such as Gly4Ser and GlyxSery (GS) and polyglycine (G) linkers.

The enzyme enables separation of the individual domains of multi-functional fusion proteins to facilitate characterization and increase the understanding of these molecules. Middle-level analysis of fusion proteins serves to both reduce the overall sample complexity and allow for domain-specific identification and monitoring of post-translational modifications.

GlySERIAS is available as a lyophilized enzyme for digestion of up to 2 mg fusion protein.

Key Characteristics

  • Unique enzyme digesting fusion proteins with flexible linkers
  • Improves resolution and reduces complexity of fusion protein analysis
  • A middle-level approach for characterization of fusion protein components

About GlySERIAS

GlySERIAS is a unique enzyme for digestion of fusion proteins designed with flexible linkers consisting of repetitive sequences of glycine, or glycine and serine, residues. The enzyme is active under native reaction conditions and enables middle-level characterization of complex fusion proteins. The linker regions are digested at several sites simultaneously, leading to separation of the previously linked components. Optimal activity occurs at 37°C, pH 7.6 and the length of the incubation can be varied depending on the desired end-products. A shorter incubation time will allow for a more complete coverage of the linker sequence whereas a longer incubation time will result in more homogeneous subunits.

GlySERIAS is cloned from phage K and is expressed in E. coli. The enzyme contains a His-tag and has a molecular weight of 18 kDa.

Unit Definition

One unit GlySERIAS digests ≥ 95% of 1 µg dulaglutide at a minimum of one site, when incubated in TBS (50 mM Tris-HCl, 150 mM NaCl, pH 7.6) at 37°C for 15 minutes.

Content and Storage

GlySERIAS is supplied lyophilized in 50 mM Tris-HCl, 150 mM NaCl, pH 7.6, with no preservatives added. Upon arrival the enzyme should be stored at -20°C. One vial contains sufficient material to digest 2 mg fusion protein.

After reconstitution, GlySERIAS is stable for at least 1 month at +4-8°C.

GlySERIAS is for R&D use only.

  • Fusion proteins with flexible GS or G linkers
  • 1 hour reaction (optimization for individual proteins may be required)
  • No reducing agents or co-factors needed
  • Flexible linkers consisting of repetitive sequences of Gly or Gly and Ser residues

Applications of GlySERIAS

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Safety Data Sheet

Certificate of Analysis

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