Review Article

Production of monoclonal antibodies (mAbs) using Chinese Hamster Ovary

(CHO) cell line
Shubham Mishra (2012A1PS484H)
Biotechnology, or use of microorganisms in food and medicinal production has been in existence for
several generation(brewery, dairy, etc.) now, though we have only recently began using more
sophisticated and complicated processes for obtaining product of our requirement on a large scale.
Animal cell culture technology for production of bio-products has now been in major utility and
development for a significant amount of time and is considered robust and established method for
mass production. In this review we are examining the process chain for production of monoclonal
antibodies (mAbs) using Chinese Hamster Ovary (CHO) as host cell.
Introduction
With increasing use and demand for protein
therapeutic treatments, use of mammalian cell line
for development for such proteins has been a thrust
area for major biotechnology research and
development centres and biopharmaceutical
companies. Though the cell line still most widely
used is that of Chinese Hamster Ovary, which was
developed first about five decades ago and is a fairly
mature method of producing experimental proteins
through gene manipulation, we have only recently
acquired expertise to use the cell culture for mass
production of desired products on large and very
large scale and making stable, safe and cost effective
product available to market in the shortest duration
of time.1
Cell culture process can be outlined into few
sophisticated sub-processes, beginning from i)
selection and generation of optimal cell line for the
desired product, which is then ii) optimized for
suitable media for processes which would facilitate
growth of the chosen cell line, which is defined and
is satisfactory to requirement on laboratory scale is
iii) moved to pilot scale to obtain feasibility of scale
up through the designed process and produce the
desired material for toxicology, stability and other
regulatory analysis. Pilot scale also determines the
number of seed train stages, inoculum growth stages,
reaction duration and provides a template for large
scale production.

In this review we shall keep our attention to

production of mAbs using CHO cell line3, which
being a mammalian cell has certain process
bottlenecks as low yield, high mutability,
requirement of complex medium and serum and
sensitivity to shear force. This review shall be
discussing the existing process flow of industrial
production of mAbs using CHO cell line and process
extensions to increase efficacy of products and
minimize bottlenecks for cost effective production.
Process Flow and Design Parameters
After the cell line engineering challenges of
selection of correct cell line for the desired product,
gene manipulation and cloning are met in the
laboratory scale, the process engineering challenges
surface as the number of clones of the product
producing host cell is extremely small and the
process is required to scale up the production from
few pg/cell/day to several kg/culture reactor/day.
The scheme of operations (Fig. 1) shows how a vial
of clone cells is scaled up for industrial production
of a desired product from them.
As already noted that mammalian cells are difficult
to work with the simple process flow shown in the
Fig. 1 has far more complicated control and
eregulation. As we move from seed train stage to
inoculum train stage there are a number of
parameters that require attention as with scale up of
size issues like, mass transfer, heat transfer, oxygen

Figure 1. Process Scheme for Cell Line Technology Scale Up

concentration and homogeneity. Apart from these,

choice of type of reactor at each stage, feed and
media constituents, feed blow if we are using a
chemostat are also very crucial to the product
development.
For CHO cell line development initial reactor types
are batch reactors as the number of cells is very low
and very high amount of control is required in
addition of feed and media, hence the reactor types
used after a new vial of CHO is taken for
development through seed train development stage
is preferentially batch reactor. Development of
media for mAbs development for mammalian host
cell has been under study for over two decades and
is now fully chemically defined as composed of
vitamins, amino acids, inorganic salts as
micronutrients, lipids and insulin like growth
factors7. Once we have reached a reasonable cell
density and number of cell we can switch to larger
chemostat which would increase our throughput
hence reducing the time taken for product
development.
The physical factors of importance such as pH
control, cooling, maintenance of Oxygen demand,
maintaining mass transfer rate of all the essential
nutrients, and cell shearing is a challenge at every
stage of development, though on small scale these
issues are easy to tackle using simple marine blade

or other gentler stirrer, and cooling water jacket

around the reactor, a medium sized ring sparger
placed below the stirrer is good enough to maintain
oxygen demand of the culture8; on large scale these
parameters are difficult to maintain and achieved by
use either using multiple blades on the same stirrer
shaft along the height of the reactor, and sparger also
accordingly releasing required gases at various
points along the height for maximum mixing and
mass transfer.4,5,6
Conclusion
Animal cell line technology is a mature and robust
method for production of a large variety of
therapeutic proteins. mAbs are also produced using
the same process which in practicality is a
complicated process with a large number of
parameters to be controlled for a long duration of
time as biological culture development take a
reasonable amount of time for growth. Issues of
mass transfer, oxygen demand, pH control,
temperature control are very crucial due sensitive
nature of the cell culture.
References
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2. Figure Reference Recombinant DNA

Technology in Emerging Modalities for
Melanoma Immunotherapy, Vitali Alexeev,
Alyson Pidich, Daria Marley Kemp and Olga
Igoucheva, Chapter 6, "Melanoma - From
Early Detection to Treatment", book edited
by Guy Huynh Thien Duc, ISBN 978-953-510961-7

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