Explore the Journey of Therapeutic Antibody Production

The therapeutic antibody journey starts
with a specific antibody gene sequence,
which is optimized and inserted into the
genome of host cells – typically Chinese
hamster ovary (CHO) cells.
This development process typically takes
6–12 months and involves numerous
analytical tests – including sequencing and
binding assays – to ensure the cells produce
antibodies with the correct sequence,
structure and functional properties.
Therapeutic antibodies represent one of the most
important advances of the modern biopharmaceutic
landscape, with over 100 approved treatments and
thousands more in development.1 These complex
proteins can precisely target disease mechanisms
while minimizing side effects, making them ideal
therapeutic tools against hard-to-treat diseases.
This infographic explores the journey involved in
therapeutic antibody production – from initial
engineering to final purification.
Engineered cells then undergo extensive
screening and characterization, with
only one candidate making it through
to become a production cell line out of
thousands of possibilities.2
The selected cells must demonstrate key
criteria, including:
Consistent
growth
Genetic
stability
High
productivity
Engineering the perfect producer
THERAPEUTIC ANTIBODY
The Development Journey of a
Initial cell culture and expansion
Harvesting the desired product
Purification and polishing
Final processing
This phase begins in small shake flasks
containing just a few hundred
milliliters of nutrient media.
These volumes are gradually increased and,
depending on manufacturing requirements,
can be carried out in production vessels
holding 10,000–20,000 liters.3
Throughout the scale-up
process, various parameters
are monitored to maintain
optimal conditions for
cell growth and antibody
production.
A combination of automated
and manual technology is
used to continuously monitor
and adjust pH, temperature,
dissolved oxygen and
nutrient levels.
This culture phase typically runs for 10–14 days,
during which cells multiply and produce antibodies
that accumulate in the culture medium.
After primary recovery, depth filtration steps remove finer particles.
Throughout this process, conditions are carefully controlled to prevent antibody degradation
or aggregation, maintaining the temperature, pH and ionic conditions within narrow ranges.
Primary recovery operations remove intact cells
and larger debris, typically using industrial
centrifuges or specialized filtration systems.
The entire process operates under aseptic
conditions to prevent contamination, with extensive
in-process testing to verify product quality.
The harvested fluid is a complex mixture containing
the target antibody along with host cell proteins, DNA
and media components. It must meet strict quality
specifications before advancing to purification.
This stage begins with capture
chromatography, typically using Protein A
resins – specialized materials that selectively
bind to the constant (Fc) region of antibodies.
This method can achieve over 95% purity in a
single step, concentrating the antibody while
removing the majority of contaminants.
This phase begins with viral filtration – specialized nanometerscale
filters that provide an additional safety measure by removing
potential viral contaminants. The antibody solution then undergoes
ultrafiltration and diafiltration to concentrate the antibodies.
The formulated drug substance is then sterile filtered
and aseptically added to vials or pre-filled syringes
using high-precision automated equipment.
When sufficient volumes of antibodies
have accumulated and reached peak
concentration, the harvesting process begins.
The purification phase transforms
a complex biological mixture into a
highly pure product through a series
of chromatography steps. Each step is
designed to separate the target antibody
from process-related impurities.
Ion exchange
Hydrophobic interaction
Size exclusion
Production process
STAGE 1
STAGE 3
Purity
>99% Purity
Sample
Target antibodies
Protein A resins
95% purity
WASTE LARGE DEBRIS
Following capture, intermediate and polishing
chromatography steps employ different separation
principles to remove remaining impurities.
The entire purification process is validated to
consistently deliver a final product meeting stringent
purity requirements: typically > 99% pure, with
impurities measured in parts per million.
Formulation is then used to combine the antibody
with carefully selected excipients –such as
buffering agents, stabilizers and tonicity modifiers
– to maintain stability, prevent aggregation and
ensure patient comfort during administration.
References
1. Lyu X, Zhao Q, Hui J, et al. The global landscape of approved antibody therapies. Antib Ther. 2022;5(4):233–257. doi: 10.1093/abt/tbac021
2. B ailly M, Mieczkowski C, Juan V, et al. Predicting antibody developability profiles through early stage discovery screening. mAbs. 2020;12(1):1743053. doi: 10.1080/19420862.2020.1743053
3. Kelley B. Industrialization of mAb production technology: The bioprocessing industry at a crossroads. mAbs. 2009;1(5):443–452. doi: https://doi.org/10.4161/mabs.1.5.9448
The entire process, from initial cell culture to final product, operates under stringent good
manufacturing practice (GMP) regulations to ensure consistent quality and patient safety.
Each batch undergoes extensive quality control testing
including identity, purity, potency, sterility and endotoxin testing,
before being labeled, packaged and released for distribution.
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Steven Gibney, PhD
Illustrated by Alisha Vroom