Hi everyone,
I'm a researcher working with proteins and I have a question about performing GPC (size exclusion chromatography).
Lysozyme is known to hydrolyze peptidoglycan, which is a component of bacterial cell walls. Given that GPC resin is composed of agarose and dextran, I'm wondering if lysozyme could interact with the GPC resin.
The molecular weight of lysozyme is approximately 16kDa. When I equilibrate a GPC column (HiLoad 16/600 Superdex 75pg) with 20mM phosphate buffer and 100mM NaCl and then load lysozyme, the retention time corresponds to about 8-9kDa. This difference in retention time makes me suspect that lysozyme is interacting with the GPC resin.
What are your thoughts on this? Could the interaction between lysozyme and the GPC resin be causing this discrepancy in retention time?
"A rapid method for the purification of lysozyme (mucopeptide N-acetyl-muramoylhydrolase, EC 3.2.1.17) from hen egg-white has been devised. It was that gel filtration chromatography on agarose columns can be used selectively to purify lysozyme, due to the fact that this protein interacts with the agarose matrix and elutes later than the corresponding total volume for the column. Thus, lysozyme is directly obtained in a relatively pure form and with a high specific activity. In principle, this simple method can be used to prepare lysozymes from other sources."
Article A simple, one-step chromatographic procedure for the purific...
Lysozyme can interact with both dextran and agarose. Research shows that lysozyme binds to Sephadex G75, a dextran-based matrix, in a pH-dependent manner. Additionally, lysozyme exhibits anomalous behavior on agarose columns, indicating an interaction between lysozyme and agarose. These interactions are important for protein research, particularly in size exclusion chromatography (GPC), where understanding these interactions is crucial for accurate protein analysis and separation.
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