Hi all,
Can the sequence of protein change during purification? (e.g substitution)
What could be the reasons? It probably depends on the type of purification used. For the context, the purification would be a "conventional" HPLC or UPLC.
Dear Sir. Concerning your issue about some reasons for sequence changes during protein purification . I think the following below links may help you in answering your reasoning:
http://wolfson.huji.ac.il/purification/PDF/ReversePhase/VYDAChandbookRPC.pdf https://www.hplc.eu/Downloads/ACE_Guide_Peptides.pdf
Thanks
Modifications can occur, such as loss of phosphates, oxidation of cysteine, and proteolysis. Lysine residues may become carboxylated in some proteins, although this is reversible and is dependent on the tertiary structure. I suppose deamidation of asparagine or glutamine is a possibility under some circumstances. There could be read-through of a stop codon. There could be a mutation introduced during preparation of the expression plasmid.
What sequence change do you think has happened?
dear Van Hh,
Except last sentence, I agree with Adam Shapiro.
Piyal Aravinna
- Badges
- Science method