I have used all three lines. I use the cells adapted to the same media for 
comparison for reduced costs (ie not the kit or recommended media they are 
normally sold with). Doing it this way the difference is not exceptional, so we 
just use normal 293E cells adapted to suspension. If you are using the supplied 
kit reagents and media, Expi293 does work exceptionally.

There is also ExpiPro293 now.

We typically find secreted proteins are better produced in CHO (ie ExpiCHO), 
and that is also the industry standard for most biologics. Your construct, 
signal peptide and codons also make a big difference.

Best

Kelvin

--
Kelvin Lau
https://people.epfl.ch/kelvin.lau

Schedule a 30 minute 
meeting<https://outlook.office.com/bookwithme/user/[email protected]?anonymous&ismsaljsauthenabled&ep=plink>


Scientist - Protein production and structure core facility – PTPSP
President – EPFL Scientific Staff Association – ELSA

EPFL SV PTECH PTPSP
AI 2146 (Bâtiment AI)
Station 19
CH-1015 Lausanne
Switzerland
Email: [email protected]<mailto:[email protected]>
Phone: +41 21 69 30267<tel:+41%2021%2069%C2%A030267>
If unreachable: +41 21 69 34494<tel:+41%2021%2069%C2%A030267>

On 11.02.2026, 16:07, "CCP4 bulletin board" <[email protected]> wrote:

Hello Everyone,

I am seeking advice on selecting the appropriate cell lines for the expression 
of a secreted protein that is over 180 kDa in size. I am particularly 
interested in conducting expression experiments using mammalian cells. I have 
come across several research papers that mention the use of HEK-derived cell 
lines, such as HEK293 GnTI-, Expi293, and FreeStyle 293F.

I would greatly appreciate your insights on how to choose the best cell line to 
achieve a high protein expression yield, as this specific protein has shown 
suboptimal expression in other systems.

Thank you for your help.

I look forward to your response.

Regards,
--
Anamika Singh, Ph.D.
Research Associate


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