Can anyone advise me on leader sequences to obtain recombinant proteins that are secreted by the cell culture? I should use mammalian cells for expression (from the Expi293 kit by ThermoFisher), and I would like my protein to be secreted. There is literature suggesting the use of the Ig-k chain or the CD33 sequence, but I can't find the reference coding sequence. Also, since the leader sequence would need to be added to a primer for PCR, is there a risk of it becoming too long?
Thank you for your help
Dear Laura
i successfully used the following leader sequences
1) MRPGTALQAVLLAVLLVGLRAATG
and
2)MVLAQGLLSMALLALCWERSLAGA
those work both quite well for antigens produced in Expi293
while i'm using the following sequence:
1) MGWSCIILFLVATATGVHS
for monoclonal antibodies produced in ExpiCHO
Regarding the addition to primers, i suggest to you to introduce it in your trasfection vector. You can easily modify it using the PIPE cloning as reported in my post:
https://proteocool.blogspot.com/2021/09/tips-of-week12-rapid.html
best regards
Manuele
