Hi, I did fractionation of crude peptides with a gradient of mobile phase A of 0.1% TFA and mobile phase B of acetonitrile+0.1% TFA. After the lyophilization of fractions, I tested their antimicrobial activity and all the fractions showed zone of inhibition in disc diffusion assay. Now I am wondering whether the effect from TFA paired peptides. My question is how can I remove the TFA from the obtained fractions to test the antimicrobial activity.
Hi Md Atikur Rahman,
The TFA is volatile and should be removed during the lyophilization step. After the lyophilization step, what do you use to re-suspend the peptides? Are you able to resolubilize the peptides successfully?
Thank you,
Hediye.
Hi Md Atikur Rahman,
I have experience where the pH of resuspended peptides can be acidic even after lyophilisation. Check the pH after you resuspend the peptides. In the past, I needed to resuspend and lyophilise a few times to ensure the pH was neutral before proceeding. In my case, I used TEAB or ammonium bicarbonate for resuspension as both are volatile.
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