Analytical characterization methods are necessary to ensure the identity, purity, structural and conformational integrity, and activity of an enzyme product. Creative Enzymes provides a broad range of latest analytical testing techniques to support enzyme characterization throughout all project stages. To meet the specific requirements, Creative Enzymes also offers bespoke enzyme characterization services, from which our customers could benefit from the complete analytical characterization packages. The expert team of Creative Enzymes, complemented by our state-of-the-art technology, supports customers with the highest standards of quality, technical analysis, and data documentation.

Enzyme characterization is one critical part of recombinant enzyme expression. Enzyme quality control technology platform of Creative Enzymes is aimed at providing professional technical support for identifying and characterizing the final enzyme products. The analysis is available for raw, intermediate, and final enzyme products through multiple characterization techniques. Our enzyme analysis services include but are not limited to the following:

• Measurement of the enzyme molecular weight
  a. SDS-PAGE: the most widely used method to separate enzyme from mixtures according to their electrophoretic mobility differences caused by the length of polypeptide chain or molecular weight.
  b. Western Blot: the identification of enzymes by their immunologic reaction with antibodies of known specificity.
  
• Assessment of the enzyme purity
  a. SDS-PAGE
  b. HPLC: to determine the identity, content, and purity of an enzyme. SEC-HPLC can be used to determine degree of aggregation.
  
• Measurement of the enzyme concentration
  a. ELISA: antibodies specific identification of an enzyme.
  b. BCA assay: using biuret reaction with BCA to yield a purple-colored product that is used to quantify the enzyme.
  c. Bradford assay: a colorimetric assay based on the interaction between Coomassie brilliant blue and arginine and aromatic residues in enzyme.
  

• Enzyme identification by mass spectrometry
  An analytical technique that sorts the ions based on their mass-to-charge ratio. The two major methods for ionization of whole enzymes are electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI).
• Detection of the endotoxin level
• Amino-terminal (N-terminal) sequence analysis
• Evaluation of enzyme stability
• Enzyme activity assay
• Enzyme steady state kinetics measurement
• Inhibitors and activators characterization
• Evaluation of substrate preference
• Evaluation of optimal activity (pH, temperature)
• Analysis of the enzyme subunit aggregation
• Analysis of the enzyme structure
• Crystallization service
  Structural analysis by X-ray diffraction, neutron diffraction, and some techniques of electron microscopy using enzyme crystals is able to determine the molecular structure of the enzyme. Multiple enzyme crystallization techniques are available, including salting out crystallization technique, organic solvent crystallization technique, dialysis balance crystallization technique, composite crystallization method, and isoelectric point crystallization technique.

Overall, the expertise of Creative Enzymes in biochemistry and enzymology offers a full enzyme characterization service to all customers. We provide a comprehensive list of characterization with different focuses, such as molecular weight and subunits, optimal activity, inhibitors and activators, substrate preference among others. With the strength in enzyme related analysis, Creative Enzymes offers both typical and bespoke tools for your most challenging technological projects. These services are designed to satisfy all the needs in research and product development.