Temperature control and CD/fluorescence spectrum of lysozyme using temperature/wavelength scan measurement program

Recently, there has been a significant increase in the research and manufacturing of biomedicines derived from proteins, which are becoming more widely available in the bio-phamaceutical industry. An important requirement in the manufacturing and quality control of protein-based biopharmaceuticals is in the assessment of stability during storage and the affects of storage conditions. The measurement of denaturation and thermal stability are of considerable importance in guaranteeing the efficacy of biopharmaceuticals. CD measurement offers significant advantage in the assessment of protein secondary structure due to its requirement for small amount of sample coupled with high sensitivity measurement. Therefore, CD measurement is becoming one of the most popular techniques used in the analysis of the thermal stability and changes in protein structure caused by ionic strength and pH. The use of fluorescence spectroscopy in the probing tryptophan residues also yields important information about the tertiary structure of proteins.

The Jasco J-1500 temperature/wavelength scan measurement program allows the measurement of a CD spectrum at a specific temperature, as well as temperature controlled measurement at a fixed wavelength. The optional FMO-522 emission monochromator also allows simultaneous measurement of a fluorescence spectrum.

In this application note, Jasco describes the temperature controlled simultaneous measurement at 222 nm of both CD and fluorescence signals for a lysozyme sample using the J-1500 CD spectrophotometer.