i am working on lysozyme protein aggregation study (effects of several chemicals on its structure), i need suggestions for the buffer medium i should consider and why?
My suggestion is to read published papers on this subject, and make a list of the various buffers people have used for this type of experiment. See if there is any particular set of conditions that is considered standard. If not, it might be an interesting investigation to see the effect of different pH and ionic strength conditions.
Sir, i have gone through literature and majority suggests Sodium phosphate buffer of different concentration but can i use PBS for this kind of studies?
Depends on how your going to follow the aggregation, for CD very low buffer concentration must being performed, 10 mM phosphate, citrate or acetate. Also depends on pI of protein or in which conditions you want to test its stability, or the amount of aggregation. Also, you can use a metal (copper, iron, nickel) I performed an aggregation test using Iron 2+, 6 micro molar at pH 4.0 (acetate 10 mM) for interferon and followed by SEC, I was successful. But it depends on protein.
PBS contains salt (NaCl and KCl) in addition to sodium and potassium phosphate. If that is going to be a problem, use the sodium phosphate alone.
PBS may be used for studying lysozyme aggregation. Please go through the article "Fibrillation of hen egg white lysozyme triggers reduction of copper(II)". There are other articles also by the group on the same topic.
It really depends on what you want to study. Lysozyme is a very robust protein and can handle different buffers, pH, conditions with ease. The presence of salt will change the aggregation behaviour quite significantly though and PBS has quite a bit of salt.
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