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Changing the Treatment Landscapewith the Development of Biologics
By Leala Thomas and Payal Khandelwal

Medicines called biologics are transforming the healthcare landscape by offering hard-to-treat patientsa real treatment benefit. As targeted therapies, they have brought us much closer to the realization ofpersonalized solutions to our health problems. A biologic or biological drug is manufactured using livingorganisms, a different process from that used for small molecule drugs, which are typically produced
via chemical synthesis. Biologics have seen a rapid rise in popularity as treatment options for many
diseases; even offering cures where none existed before. Why is this true? What do biological drugs
offer that other therapies do not?
Novel Therapeutics Called Biologics
Biologics are not a new class of drugs, as biological drugs suchas insulin have been available for decades. What changed isthe availability and potential applications for biological drugs.
Our increased understanding of the biological processes thatare part of human health and the changes that occur during
disease or chronic illness have helped spur the development ofnovel biological drugs targeted to provide treatment and relief
for patients (Morrow and Felcone 2004). Biologics are large,
complex molecules consisting of proteins, peptides, nucleic
acids, sugars, other cellular structures, or a combination ofthese, produced within living cells or microorganisms. The
shifting focus toward biological drug products results intargeted treatments that have mechanisms of action previously
unavailable from existing small molecule drugs.
Both small and large molecule drugs interact with a patient’s
biology; however, small molecule drugs work as inhibitors
disrupting a process ordinarily associated with a particulardisease. As small molecules that can penetrate cells, thepotential exists for off-target interactions that cause a myriadof side effects. Large biologic drugs, in contrast, are designed
to bind to specific targets with extreme precision. Sometimes

they can stimulate the immune system to react to thepresence of a problem such as a tumor. Biologics target
molecular processes that small molecule drugs cannot.
As healthcare requirements around the world have evolved
due to the rise in aging populations and an increase incountries prioritizing the health of their citizens, researchfunding for diseases and treatment options has also
accelerated. Recently developed biological drugs, for example
Abbvie’s HUMIRA and Roche’s RITUXAN, play a critical rolein the management of many serious illnesses, including rare
genetic disorders, autoimmune diseases, and cancer(Figure 1; Gottleib 2018). Often they are the only available
treatment option for the patient.
In terms of approvals for novel medicines, biological drugshave grown to comprise approximately one-third of all new
medicines approved by the U.S. FDA (Gottleib 2018). In
addition, more than half the drugs currently in developmentby pharma and biotech companies are biologics. Cancer
research receives the most R&D funding for developmentof novel biological drugs with rare diseases, autoimmunedisorders, and neurological disorders occupying the next
three spots (Hooven 2017).

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Cancer

978

Orphan diseases

465

Inflammatory/immunological diseases

317

Central nervous system diseases

224

Blood diseases

205Diabetes/related conditions

143

Heart disease and stroke

113

Alzheimer’s dsease 420100200300

400500600
Number of products

700800900

1,000Fig. 1. Therapeutic focus of the biologics in clinical development. Adapted from Hooven 2017.

Focus on Monoclonal Antibodies
Monoclonal antibody (mAb) biological drugs top the list whencompared to other biological drug types, such as proteins,enzymes, and vaccines. In addition to an increase in spending
on R&D for biological drugs, supportive government initiatives
and demand for personalized medicines are contributing tothe expectation that the global market for mAbs will be worth$138 billion by 2024 (Grand View Research 2016).
Manufacturing Monoclonal Antibodies
In development of a biological product, the resulting productis dependent upon the manufacturing process, which is time
consuming, challenging, expensive, and often complicated. In
addition to the usual clinical trials required to establish safety
and efficacy, development includes a complex manufacturing
process that includes genetically engineering a cell to produce
a protein, testing to ensure quality, and harvesting, purifying,and stabilizing the protein (Jallal 2017).
The process of manufacturing a fragile, sensitive biologicalmolecule, such as an antibody made from living cells, hascomplex requirements that include fermentation, aseptic
processing and purification, storage, and testing. The active
component of the drug, often a portion or modification of theoriginal protein or polypeptide, may not be clearly characterized.
Due to the heterogeneous nature of the molecule, the biologic
can have an impurity profile that varies with the manufacturing
and testing process utilized. One of the primary confounding
issues is the need to identify a purification strategy thatproduces the highest yield while maintaining the purity of
the final product. This critical step in the process is often
accompanied by very significant costs.
Recent advances in upstream processes have improved mAb
titers in mammalian cell culture. Increases in the fermentation
volume and protein mass have increased the challenges
involved in processing harvested material. As purification
processes have improved and yields have grown, the incidenceof impurities has also grown and had a negative impact on thepurity of the final product. In addition, elevated process andproduct related impurities from prolonged fermentation andhigher cell density are further complications (He et al. 2011).
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For downstream mAb purification, column chromatography
is normally preferred, using a two- to three-step process that
includes capture, intermediate, and polish steps (Figure 2) andselecting specific resins based on their technical parameters.
But not all resins are created equal. The ideal strategy canbe customized according to final intended use and include
considerations for cost, harsh elution conditions, desired
monomer recovery, and purity.

UNOsphere
SUPrA Resin

UNOsphere Q Resin

CHT Ceramic
Hydroxyapatite

CaptureIntermediate polish

Final polishmAb1 (chimeric IgG1)
HCP = 175 ng/mg
DNA = 18.6 ng/ mg
Protein A = 3 ng/mg

HCP < 3 ng/mg
DNA = 3.5 ng/mg
Protein A = 0.8 ng/mg

HCP < 2 ng/mg
DNA = 2.5 ng/mg
Protein A < 0.7 ng/mg
Aggregate < 0.03%

Fig. 2. Example of a mAb purification workflow.

Protein A chromatography has been the method of choicefor the initial step of purification because it is relatively easy
to use without modification and has the ability to remove
host cell proteins, nucleic acids, endotoxins, and viruses.
However, Protein A is an immunotoxin and capable of forming
neutralizing antibodies (Gagnon et al. 2006). Protein A is
also quite expensive. An ion exchange resin such as Nuvia S
provides an economical and effective alternative to Protein A
for the capture step. Nuvia S is an ultra-high capacity cationexchange (CEX) resin that provides high dynamic binding at abroad range of pH and conductivity with low backpressure athigh flow rates. High flow rates minimize the mAb’s exposure
to the proteases and nucleases that exist in cell culture.
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Nuvia HR-S, another high-resolution CEX resin, can be used
for both the intermediate and polish purification steps. It
is ideal for separation of closely related biomolecules andimpurities like mAb aggregates (Khandelwal 2016). Nuvia
cPrime, a hydrophobic CEX resin, has unique selectivity thatuses hydrophobic and CEX interaction modes to achieveeffective purification in the intermediate and final polishsteps. Its particle size is optimized to deliver exceptional flow
properties, fast mass transfer, and stability.

References

The gold standard for aggregate removal is CHT Ceramic
Hydroxyapatite, a highly effective mixed-mode media. CHTremoves aggregates from mAbs without affecting purity or
yield of monomer mAbs (He 2015). CHT binds to biomoleculesby calcium metal affinity, phosphoryl cation exchangeinteractions, and hydrogen bonding.

He X (2015). Mixed-mode chromatography for mAb S aggregate removal:
Comparison of CHT Ceramic Hydroxyapatite, Capto adhere, and Capto adhere
ImpRes. Bio-Rad Bulletin 6749.

Conclusion
Biologics are becoming the primary focus for a growing
number of therapeutic developers. Although they are bothchallenging and expensive to develop, they have applications
for a wide range of diseases that are not being adequately
addressed with existing medicines. In addition, biologics arechallenging to copy since they are produced in living cells.
As a result, the effort devoted to developing a successful
biologic is paying off. The success of a mAb in particulardepends upon its purification strategy and the effectivenessof downstream removal of aggregates.

Khandelwal P (2016). Practical guide: selecting optimal resins for monoclonal
antibody process purification. Bio-Rad Bulletin 6875.

Gagnon P et al. (2006). A ceramic hydroxyapatite–based purification platform:
simultaneous removal of leached Protein A, aggregates, DNA, and endotoxinsfrom mAbs. Bio-Rad Bulletin RP0033.
Gottleib S (2018). Capturing the benefits of competition for patients. fda.gov/NewsEvents/Speeches/ucm599833.htm, accessed December 11, 2018.
Grand View Research (2016). Monoclonal antibodies (mAbs) market size
worth $138.6 billion by 2024. grandviewresearch.com/press-release/globalmonoclonal-antibodies-market, accessed December 11, 2018.
He X et al. (2011). A purification strategy for clinical-grade monoclonal antibody
using hydrophobic cation exchange chromatography. Bio-Rad Bulletin 6241.

Hooven MD (2017). Opportunities and challenges in biologic drug delivery.
americanpharmaceuticalreview.com/Featured-Articles/345540-Opportunitiesand-Challenges-in-Biologic-Drug-Delivery/, accessed December 11, 2018.
Jallal B (2017). Realizing the promise of biologics. hhpronline.org/articles/2017/4/9/realizing-the-promise-of-biologics, accessed January 4, 2019.

Morrow T and Felcone LH (2004). Defining the difference: what makes biologics
unique. Biotechnol Healthc 1, 24–29.
Bio-Rad and CHT are trademarks of Bio-Rad Laboratories, Inc.
in certain jurisdictions. All trademarks used herein are the propertyof their respective owner.

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