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Enzymatic Reagent

Enzymatic Reagent

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Modality

Enzymatic Reagent

Tag Removal Proteases

To improve the solubility and simplify the purification process of recombinant proteins, researchers usually add fusion tags, of which His-tag, maltose-binding protein (MBP), and glutathione-S-transferase (GST) are commonly used.

While tagged as an extra sequence of protein, it should be removed to maintain biological activity in the pharmaceutical industry. The removal of fusion tags requires site-specific proteases, such as enterokinase (EK), thrombin, tobacco etch virus protease (TEVp), human rhinovirus protease 3C (HRV3C), small ubiquitin modifying protein (SUMO) protease, tobacco vein mottling virus (TVMV) protease, and carboxypeptidase A/B (CPA/CPB).

Type

Enzyme

Recognition Site

Tag removal endoproteases

Enterokinase (EK), Enteropeptidase

DDDDK↓

Thrombin

LVPR↓GS

TEV protease

ENLYFQ↓G

HRV3C protease

LEVLFQ↓GP

SUMO protease

SUMO tertiary structure

TVMV protease

ETVRFQG↓S

Tag removal exoproteases

Carboxypeptidase A (CPA)

C-terminal amino acids, except Pro, Lys and Arg

Carboxypeptidase B (CPB)

C-terminal Lys and Arg

Other Proteases

Application

Enzyme

Function

Other proteases

Protease K

A serine protease that digests proteins by hydrolyzing peptide bonds.

IdeS, IgG protease

IgG degrading enzymes (Ides) that cleave at a specific site of immunoglobulin G (IgG), generating Fab and Fc fragments

IgA1 protease

A proteolytic enzyme that cleaves specific site in the human immunoglobulin A1 (IgA1) hinge region sequence.

Nuclease

Application

Enzyme

Function

Nuclease

Nuclease

Cleaves nucleic acids (DNA or RNA) by hydrolyzing the phosphodiester bonds.

Restrict enzyme

An endonuclease that cleaves DNA at or near specific sites.

Amidase

Application

Enzyme

Function

Amidase

PNGase F

Cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides.

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