Scale-up recombinant protein production from HEK or CHO cells

Produce high yields of recombinant protein from stably-expressing mammalian lines

A compact FiberCell hollow fibre bioreactor is ideal for producing from 50 mg up to 1 gram of protein from stably transfected adherent or suspension cells. High density conditions free of shear stress strongly favour the complete and uniform glycosylation of secreted proteins. Examples of successful projects include: “difficult-to-express” multi-subunit cytokines or recombinant multiple subunit vaccines, fusion proteins and humanized mAbs. See : publications.

A FiberCell bioreactor enables you to keep total control of your production process in a standard cell culture lab and requires just the space of an incubator shelf. Find out more about hollow fibre bioreactor technology here.

“…the high yields of pure IL-12 obtained using the method described in this study will provide an avenue to embark on structural studies aimed at the development of novel IL-12 based therapeutics for the treatment of cancers and infectious diseases.”

See publication

Fits on shelf of incubator

Continuously culture more than 1E+9 cells in a 20ml Medium Size FiberCell Cartridge Module and routinely harvest concentrated supernatant. A FiberCell bioreactor with either one or two independent cartridge modules fits easily on one shelf of a standard 180L lab incubator. Choose between 20 ml or 70 ml cartridges with maximum productivity equivalent to daily harvests from 200 roller bottles.

  • Concentration typically 50 times higher than flask or rollers
  • Efficient production of “difficult-to-express” recombinant proteins
  • Simple protein-free media […learn about serum free operation]

The hassle of working with flasks or roller bottles can be avoided

The idea of scaling up a mammalian cell culture process may be daunting. However, passaging cells in large numbers of flasks or roller bottles can easily be avoided. Continuous high-cell-density culture using a compact FiberCell hollow fibre bioreactor enables any cell culture laboratory to keep control of a production process without recourse to outsourcing.

Check what your scale-up factor could be

Quickly calculate the amount of recombinant protein you could produce per month with your clone: [CALCULATE]

Customer Publications:

See our customer publications

Example : failure in flask culture, success in hollow fibre

A complex hexamerized IgG consisting of six IgG1 subunits held together with three IgA tails was produced in flask and FiberCell culture.

In flasks approximately 40% of the protein was secreted as an unfolded monomeric subunit whereas >90% was correctly folded in FiberCell culture.

476 mg of protein was produced in two months in a harvest volume of less than 5 L using FiberCell cartridge C2018. (Data courtesy of Dr. Jim Arthos, Bethesda, MD.)

Examples of challenging applications

Obtain well-folded and glycosylated recombinant protein

The high density, zero-shear stress, constant pH culture environment of a FiberCell bioreactor is ideal for generating significant amounts of highly pure, correctly folded and fully glycosylated recombinant protein.

Do you need 500 mg or more?

CHO cells growing in hollow fibre bioreactor

70 ml cartridge modules for 5 x 10e10 cells

FiberCell C2003 5 Kd and C2018 20 Kd cartridge modules enable you to maintain huge numbers of cells with little maintenance and are recommended for projects to produce >=500 mg of recombinant protein. FiberCell hollow fibre cartridges typically produce around 100µg/ml/day. See cartridge modules page.

Duet pumps maintaining approx. 4E+11 cells

The compact hollow fibre culture set-up on the right is producing around 2 grams per month of a highly valuable cytokine. The large autoclavable carboys minimize hands on time. Result: a highly productive, easily affordable and low maintenance system running free of serum or extraneous media proteins.

Large FiberCell Hollow Fiber Cartridge Module

Recombinant IgG production example

Production of a Recombinant IgG in 293 Cells Using FiberCell Systems CDM-HD (Serum-Free Medium)

A medium sized 20 ml C2011 cartridge module was used to produce 276 mg of protein in a total volume of 900 mL over a 2 month period using serum-free medium. Bioreactor media was not changed at weekends but this had little overall impact on productivity.

Click to View Image

“An ideal system for generating bi-specific antibodies (bsAbs) for preclinical studies”

3D high density culture conditions

Continuous high cell density conditions

Maintain HEK293 or CHO cells at highest possible cell density (>10e8 cells per ml) without shear stress. Cells proliferate around the fibres in a more physiologic in vivo-like environment compared to flask culture and are not subjected to passaging since cell mass is gently reduced during the routine harvesting procedure.  Culture conditions in this system are ideal for superior protein quality and biological activity. You should be able to count on harvests of 2 mg to 10 mg per day of fully biologically active protein, depending on the cartridge size used, see : cartridge modules.

Cells lose dependence on serum

Under the high density conditions described above cells can be maintained free of serum in the media. In fact, once the culture is established it is generally possible to make a single-step switch to a low-cost, simplified DMEM-based media. Several examples are provided below. FiberCell provides a popular serum-substitute specifically formulated for the bioreactor, see: CDM-HD.

Adaptation from DMEM-FBS to serum-free medium

SDS-Page gel of successive harvests of a secreted recombinant protein from a 293 cell line in the FiberCell HF Bioreactor. Starting media is DMEM with 10% FBS (Monday, left). Shows a one-step switch to serum free medium (Hyclone) while maintaining high levels of the secreted target protein.

Gel showing purity of target recombinant protein

Switch from FBS to CDM-HD serum replacement (293T]

C2011 cartridge was used to produce rec. IgG with 293t cells. Note the reduction in contaminating serum proteins after switching to CDM-HD. By harvesting every day elimination of serum proteins would be even faster than shown. Note how the IgG becomes the dominant protein in the unpurified supernatant.

Switch to CDM-HD serum replacement (BHK]
BHK cells expressing PEDF using DMEM 10% FBS followed by a switch to DMEM 10% CDM-HD on day 17 with no adaptation.

Data demonstrates stable glucose uptake rate and no loss of PEDF production when using CDM-HD.

BHK cells can be cultured in hollow fibre bioreactors using protein free medium with no loss of production.

Cartridge usedCat # C2008
Cells inoculated3 X 108 cells
Medium Consumed14 L
Total protein produced99.8 mg
Total volume of harvest164 mL
Average concentration0.6 mg/mL


* indicates required
I'm interested in scaling up production of:
Send updates about:
My lab grows:

Request Quotation