My protein after column purification performing buffer exchange at 10kDa membrane with formulation buffer (50mM PB+ 100mM NaCl + 0.25mM EDTA, pH 7.3 & Cond.: 16mS/cm). After buffer exchange i am concentrating up to 15mg/mL. But retentate material is precipitating and thread like particles are forming. Can anyone suggest me is their any buffers or additives can control or stop the precipitation or aggregation.
I agree with the comments given by Vicente. Do you need too high concentration of the protein? You can use a high-throughput screening platforms for identification of your formulation buffer conditions that can maintain your proteins in soluble and stable forms. Here is the paper for your reference to use DLS-based high-throughput screening techniques for buffer additives.
https://www.researchgate.net/publication/275413441_A_rapid_and_simple_screening_method_to_identify_conditions_for_enhanced_stability_of_modular_vaccine_candidates
Article A rapid and simple screening method to identify conditions f...
Hi Nagabushan,
Before going for 15 mg/ml concentration, please do three dia with the same buffer at three stages like 5, 10 and 15 mg/ml.
Also look at the temperature when you do concentration - it depends upon the protein. But usually below 8'C deg should be good.
Before going for UF, Perform throrough 0.7% CIP and wash it so that left over residues will be cleared from your membrane/fiber which will create major issues during concnetration.
Neutralize with the buffer and then start with your product.
Check your protien for conductivity, if your 16ms/cm is very high then reduce and match with your protein product.
regards
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