I have tried to overexpress and purify a membrane protein in HEK293T by PiggyBac system with PA-tag. However, the protein expression level is very weak, so it is difficult to purify this protein with anti-PA sepharose beads when protein untarget contaminants go through column with target proteins in elution buffer. Anyone know how to enhance expression of membrane protein and avoid protein contaminants? Thank you so much!
Study of membrane glycoprotein is very important to clarify the brain function in humans.
Therefore, we have studied biotinidase and lipoamidase of brain (please see files; Brain-BIN pig and Purify brain lipoamidase).
Article Over-expression of secreted proteins from mammalian cell lines
Hi Han,
I wonder if you have solved the problem you encountered in the experiment. If the cell state, expression conditions and other factors are excluded (because the mammalian cell expression system is already very good), can you consider doing truncation? Membrane protein expression and purification is indeed a challenge, I know Creative Biostructure has a membrane protein platform, I feel it is pretty good, you can try. https://www.creative-biostructure.com/mempro%E2%84%A2-membrane-protein-platform-447.htm
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