We are now working for already one year to express our protein of interest - it is highly glycosylated, and expression does not work at all in e. coli. so we switched to a baculovirus system in sf9 insect cells. our construct seems to be okay, transfection & virus titer amplification works, but our protein (not secreted) is not really expressed. we do get some slight bands from time to time, but it's rather faulty.
does anyone have any hints how to accomplish proper expression?