Protein Aggregation During Purification

Biochemistry
1

Hello People

My recombinant protein looks fine initially but starts forming aggregates during purification and storage. Which parameters should I check first—pH, salt concentration, temperature, or protein concentration?

Thanks

2

Hi

I faced a similar issue with my recombinant protein during my PhD work. In my case, aggregation increased mainly at higher protein concentrations. I would first try keeping the protein at 4°C and testing a few pH and salt conditions. Also, check the protein immediately after purification and again after storage using SDS-PAGE or SEC. That can help identify when the aggregation is starting.

I hope it helps.