Open Access Peer-reviewed Research Article

Determining the degree of denaturation of bovine serum albumin using a new UV analysis technique

Main Article Content

Pooya Afaghi
Michael Anthony Lapolla
Khashayar Ghandi corresponding author


There is a lack of fast and inexpensive analysis methods to study the conformational changes and the degree of denaturation of proteins quantitatively. As such, a novel analytical technique is developed based on the ultraviolet-visible (UV-Vis) absorption spectrum of proteins, and a mathematical modeling of the results. The phenomenon behind this technique is the shift of the absorption peak of amino acid residues of BSA such as tyrosine, phenylalanine, and tryptophan as the protein unfolds and these residues are exposed to the solvent. However, the portion of the peak that is shifted is miniscule and it can be enhanced by using the proposed technique in this paper. As an example, we also show how this technique was applied for evaluating the temperature effects on thermal denaturation of bovine serum albumin (BSA) protein. A degree of denaturation curve as a function of time was obtained at three different temperatures using this technique. The results are reproducible and consistent with those reported in the literature. This technique is especially recommended for analyses where several tests are needed quickly, and the amount of sample is limited. Among the applications, it can be used for evaluation of disinfection through assessing the degree of denaturation for pathogens proteins.

UV-vis spectroscopy, protein denaturation, bovine serum albumin, tyrosine, phenylalanine, tryptophan

Article Details

Supporting Agencies
This work was supported by Department of National Defence of Canada through the Innovation for Defence Excellence and Security (IDEaS) program.
How to Cite
Afaghi, P., Lapolla, M., & Ghandi, K. (2021). Determining the degree of denaturation of bovine serum albumin using a new UV analysis technique. Chemical Reports, 3(1), 173-176.


  1. Telser A. Molecular Biology of the Cell, 4th Edition. Shock, 2002, 18: 289.
  2. Wrapp D, Wang N, Corbett KS, et al. Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science, 2020, 367(6483): 1260-1263.
  3. Nguyen HL, Lan PD, Thai NQ, et al. Does SARS-CoV-2 Bind to Human ACE2 Stronger Than SARS-CoV? The Journal of Physical Chemistry B, 2020, 124: 7336-7347.
  4. Wang Q, Zhang Y, Wu L, et al. Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2. Cell, 2020, 181(4): 894-904.
  5. Greenfield NJ. Using circular dichroism spectra to estimate protein secondary structure. Nature Protocol, 2007, 1: 2876-2890.
  6. Matsuo K, Sakurada Y, Yonehara R, et al. Secondary-structure analysis of denatured proteins by vacuum-ultraviolet circular dichroism spectroscopy. Biophysical Journal, 2007, 92(11): 4088-4096.
  7. Antosiewicz JM and Shugar D. UV–Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 2: selected applications. Biophysical Reviews, 2016, 8(2): 163-177.
  8. Pinho Melo E, Aires-Barros MR, Costa SMB, et al. Thermal unfolding of proteins at high pH range studied by UV absorbance. Journal of Biochemical and Biophysical Methods, 1997, 34(1): 45-59.
  9. Peters T. Serum Albumin. Advances in Protein Chemistry, 1985, 37: 161-245.
  10. Mach H and Middaugh CR. Simultaneous Monitoring of the Environment of Tryptophan, Tyrosine, and Phenylalanine Residues in Proteins by Near-Ultraviolet Second-Derivative Spectroscopy. Analytical Biochemistry, 1994, 222(2): 323-331.
  11. Berger I and Schaffitzel C. The SARS-CoV-2 spike protein: balancing stability and infectivity. Cell Research, 2020, 30: 1059-1060.