Circular dichroism (CD) spectroscopy is a fast, simple, and accurate method for studying protein structures in dilute solutions. The structural analysis was performed using the circular dichroism of proteins and the difference in absorption of left and right circularly polarized light by asymmetric molecules.
The CD spectrum of the far ultraviolet region mainly reflects the circular dichroism of peptide bonds. In the regular secondary structure of proteins or peptides, peptide bonds are highly regularly arranged. The orientation of its arrangement determines the splitting of the peptide bond energy level transition. The positions and absorption strengths of the CD bands generated by proteins or peptides with different secondary structures are different. Therefore, the information of the secondary structure of the protein or polypeptide chain can be obtained according to the far UV CD spectrum of the measured protein or polypeptide, thus revealing the secondary structure of the protein or polypeptide.
All new biotechnology or biological products require spectral analysis as proof of conformity. As a reliable partner of biopharmaceutical companies, Creative Proteomics can provide customers with far UV circular dichroism spectroscopy analysis services based on GLP / GMP. The detectable molecules range from small chiral drugs and transition metal complexes to biological macromolecular structures such as proteins, glycoproteins and nucleic acids.
We Can Provide but Not Limited to:
- Determine the conformational change of protein under renaturation and acid-base conditions and even the affinity of its interaction with ligand
- Determination of changes in the secondary structure of a protein or polypeptide caused by kinetics or thermodynamics.
- Obtain data on protein folding and conformation studies (secondary structure)
- Determination of the purity of optically active substances
- Preliminary detection of target protein before crystal growth or NMR analysis
- Quantitative analysis of drugs
The Workflow of Far UV Circular Dichroism Spectroscopy Analysis
Advantages of Far UV Circular Dichroism Spectroscopy Analysis
- We have developed unique algorithms that can determine the secondary structure of proteins in greater detail and more reliably.
- Rapid turnaround time: 5-7 days to provide comprehensive report.
- Customized service: We can customize exclusive solutions for you based on your samples.
The purity of samples for CD spectroscopy must be at least 95% (HPLC, MS or SDS-PAGE). Sample concentration> 0.5mg / ml. Sample volume> 200 μg.
The buffer should not have a high absorbance in region 190-250 nm of the spectrum. For example, high concentrations of DTT, histidine, or imidazole, cannot be used in the far-UV region. For many formulated protein samples, the absorbance of the excipients prevents collecting spectra below 200 nm. If this does happen, the calculated percentage of different kinds of secondary structure will not be reliable.
Creative Proteomics' analytical scientists can provide a clear and concise written report of the spectrum data of the far ultraviolet circular dichroism analysis to help customers analyze the secondary structure of the protein. We can also provide you with a one-stop service from sample preparation, purity analysis to circular dichroism analysis.
- Miles A J, Wallace B A. Biopharmaceutical applications of protein characterisation by circular dichroism spectroscopy//Biophysical Characterization of Proteins in Developing Biopharmaceuticals. Elsevier, 2020: 123-152.