Research Article

Received: 9 November 2020 Revised: 24 January 2021 Accepted article published: 31 January 2021 Published online in Wiley Online Library:

(wileyonlinelibrary.com) DOI 10.1002/jctb.6686

Contribution of protein A step towards cost

of goods for continuous production of
monoclonal antibody therapeutics
Vikrant Bansode, Paridhi Gupta, Nikhil Kateja and Anurag S Rathore*

Abstract
BACKGROUND: Monoclonal antibodies (mAbs) presently dominate the biotherapeutic market. However, biotherapeutics in
general, and mAbs in particular, are expensive therapies and outside the realm of affordability for most of the world. In an
attempt to lower their cost of manufacturing, a novel in-house non-protein A continuous platform has recently been reported
for mAb purification. In this paper, we perform an in-depth evaluation of the process economics to assess the impact of remov-
ing the protein A step on the cost of goods (COGs).
RESULTS: The non-protein A platform was found to result in significant savings in COGs at about 23% when comparing both
platforms in batch mode. Conversion of the non-protein A platform from batch to continuous mode led to a further increase
in these savings with about 39% gains in COGs over the conventional protein A platform.
CONCLUSIONS: The non-protein A purification platform offers a viable alternative to the standard protein A platform and can
be considered when setting up a new facility. The continuous non-protein A platform can especially be considered given the
flexibility of the setup and the utility at both small and large scales.
© 2021 Society of Chemical Industry

Supporting information may be found in the online version of this article.

Keywords: continuous processing; economic analysis; cost evaluation; biotherapeutics; monoclonal antibodies; non-protein a
purification

INTRODUCTION
Monoclonal antibodies (mAbs) have been dominating and will con-
tinue to dominate the biosimilar market. They have been success-
fully used for the treatment of cancers and autoimmune diseases.
There are a growing number of mAbs being developed for infectious
and neglected diseases, including the newly discovered SARS-CoV-2
virus as well as other viral diseases like Zika and Ebola.1–6 Although
there is a steady increase in the demand of mAb therapeutics, the
manufacturing cost remains high.7–9 The raw material cost for bio-
pharmaceutical production is 20–100 times more than that for tradi-
tional pharmaceutical drugs. Biopharmaceuticals are also required in
higher doses, which further results in higher costs of treatment.6
Countries with some form of universal healthcare are better off as
the costs of biopharmaceuticals are either completely or partially
shared by the state. Patients in countries with no such facility incur
the total cost of biopharmaceutical drugs.10 Outsourcing drug devel-
opment is a strategy used by companies to reduce the development
time but this results in a higher development cost.11 The mAb mar-
ket is predicted to be around $319 billion by 2026. Even the Indian
biosimilar market is estimated to be at $6 billion by 2024.1 Costs of
cell culture media and chromatography media are two of the major
contributors to mAb production.10–13
The purification regime used for mAbs is more or less same across
the industry and can be termed as the protein A platform. Protein A
is a type of affinity chromatography which uses immobilized protein
A ligands which specifically bind to the Fc domain of mAbs, thus selec-
tively purifying the molecules from a pool of host cell proteins (HCP)/
host cell DNA (HCD).14,15 Protein A capture is generally preceded by
several clarification steps, followed by viral inactivation and additional
polishing chromatography and formulation steps.16-18 Economic anal-
ysis for the conventional protein A-based purification platform has
been performed previously by our group.19 Although the platform
has been time tested, the cost of protein A resin is almost two times
the cost of other resins. This makes protein A chromatography the
most expensive unit operation in the purification platform.20–22 Even
though robust protein A resins have been developed for higher toler-
ance of cleaning-in-place agents like NaOH, protein A ligand leaching
remains a problem for manufacturers.23 This also reduces the number
of times the resin can be reused and incurs an additional cost of anal-
ysis for protein A leachate.24
Several attempts have been made to replace the conventional
purification platform by proposing alternatives to the protein A
capture step. Researchers have developed a platform with
* Correspondence to: AS Rathore, Department of Chemical Engineering,
Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India.
E-mail: asrathore@biotechcmz.com
Department of Chemical Engineering, Indian Institute of Technology Delhi,
New Delhi, India
1

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phenylboronic acid multimodal chromatography as the capture
step followed by ion exchange chromatography and hydrophobic
interaction chromatography as the polishing steps.25 The invest-
ment was similar to that of the conventional platform while a
20% decrease in the annual operating expenditure (OpEX) was
estimated. Other researchers have evaluated several sets of chro-
matography steps as replacement of protein A chromatography
in an attempt to reduce the purification cost of mAbs.26,27
Another study explored the use of activated carbon in flow-
through mode to clear impurities and replace protein A capture.28
While the results showed product quality comparable to that of
the standard protein A platform, the researchers did not evaluate
process economics.
A non-protein A continuous processing platform for mAb purifica-
tion was reported recently.29 The platform employs a novel coiled
flow inversion reactor (CFIR) to aid in clarification after precipitation
for removal of HCP/HCD and for inactivation of enveloped viruses.
Cation exchange chromatography is used to capture the product
as well as control charge variants. Unlike protein A chromatography,
cation exchange utilizes the difference in charge of the product and
impurities at a particular pH and conductivity values for separation.
In this paper, we examine the impact of using this platform on the
economics of commercial production of mAbs.
EXPERIMENTAL
Economic analysis
The approach for economic assessment was same for both protein-
A-based and non-protein-A-purification strategies. SuperPro
Designer (Intelligen Inc., Scotch Plains, NJ) was used to make the pro-
cess flow diagrams (PFDs) of batch as well continuous manufactur-
ing mode. Figures 1 and 2 describe the PFDs for protein A and
non-protein A purification strategies as derived from SuperPro
Designer software. Figure 3 provides a breakdown of the overall
expenditure. After incorporating all the process assumptions, eco-
nomic analysis assumptions were also added to the PFDs. The eco-
nomic evaluation reports hence generated gave total capital
investment (CapEX) and OpEX for the end-to-end PFDs, along with
the unit-operation-wise distribution (discussed in the following sub-
section). The obtained data were thereafter used for calculating eco-
nomic indicators such as cost of goods sold (COGs), net present
value (NPV) and payback time. Subsequently, sensitivity analysis with
scale of manufacturing was also performed at two scales – small
scale and large scale of production. Small scale represents clinical
material generation and large scale represents a typical manufactur-
ing scale for a ‘blockbuster’ drug. Throughputs of 15 kg per annum
  Total capital investment=years of operation ð$ yr−1 Þ +annual operating cost ð$ yr−1 Þ
COGs $ g−1 =
Annual production rate ðg yr−1 Þ
and 500 kg per annum were considered for small and large scale,
respectively.19 Process parameters were optimized in house at the
laboratory scale to be used for this study.
Economic parameters and assumptions
CapEX is the same as total capital investment (TCI), as derived from
the SuperPro Designer software. TCI is derived as per Eqn (1) below:
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TCI =total plant direct cost ðTPDCÞ
+total plant indirect cost ðTPICÞ
ð1Þ
+contractor0 s fee and contingency + other cost
TPDC, TPIC and OpEX have been defined similar to our previous
work.19
All the components of CapEX were obtained by multiplying a
suitable multiplier to the equipment purchase cost, as given in
Table S1 (supporting information). However, when prototype
devices were used in PFDs, estimated assumptions and extrapola-
tions were applied, as provided in Table S2 (supporting informa-
tion). Working capital is estimated to cover expenses for 30 days
of labor, 30 days of raw materials, 30 days of utilities and 30 days
of waste treatment.
For calculating various components of OpEX, suitable
assumptions were made. Cost of raw materials was taken from
industry-specific vendor sources. Labor and facility costs were
calculated in SuperPro Designer software using Eqns (2) and
(3), as given below. Consumable costs include annual con-
sumption of chromatography resins and filters used in depth
filtration, clarification, ultrafiltration and diafiltration. Costs of
consumables were also taken from vendor sources (Table S3,
supporting information). Wastewater treatment/disposal cost
was assumed as $0.05 kg−1. Utilities include cost of air at
0.5 VVM, cost of power and cost of steam or chilled water or
glycol, depending on the use.
!
1+ benefits+ supervision
Labour cost = ðbasic rateÞ×
+operating supplies +administration
ð2Þ
×ðlabour hoursÞ
Facility cost =maintenance+depreciation +insurance
ð3Þ
+local tax +factory expenses
Various economic indicators, namely COGs, NPV and payback
time, were calculated using the SuperPro Designer software.
COGs ($ g−1) allows us to understand the cost incurred in
manufacturing per gram of the final product produced. COGs
incorporate overall CapEX incurred for an assumed 15 years of
production and annual OpEX, along with annual throughput of
final product. The equation for calculation of COGs is given below:
… ð4Þ
In order to estimate the profitability of investment in protein A
versus non-protein A purification strategies, NPV analysis was per-
formed based on 15 years of plant operation in both cases. In the
NPV analysis, 3 years of manufacturing time for plant was
assumed before the operation period. Discount rate of 7% per
annum and income tax at 10% per annum were used for the
NPV assessment. In order to calculate annual revenue generated,
J Chem Technol Biotechnol 2021
Contribution of protein A step towards cost of goods
Figure 1. PFDs for protein A platform in (a) batch mode and (b) continuous mode.
the selling price of mAbs was taken at $20 mg−1 for both protein
A and non-protein A strategies of manufacturing.
The payback time in number of years was calculated using
Eqn (5) as follows:
Net profit ð$Þ
Payback time =
Revenue ð$ yr−1 Þ−annual operating cost ð$ yr−1 Þ
Revenue and net profit (for any year) were calculated using
Eqns (6)–(8):
   
Revenue $ yr−1 =Amount of product g yr−1
 
×average price realized $ g−1 …
Net profit ð$Þ =Gross profit ð$Þ−income tax ð$Þ…
Gross profit ð$Þ= Revenue ð$Þ−annual operating cost ð$Þ… ð8Þ
Protein A platform
For the comparison of the two platforms, an IgG1 antibody was
considered as the reference mAb. The mAb had a molecular
weight of 150 kDa and a pI value of 8.7. A standard protein A
mAb purification platform can be broken down into: cell culture,
clarification, protein A capture, viral inactivation, polishing steps
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and ultrafiltration–diafiltration. The protein A platform used in this
study in batch operation consisted of a fed-batch cell culture pro-
cess in upstream using CHO (Chinese hamster ovary) cell line. The
purification regime started with harvest clarification using centri-
fuge and depth filters followed by a protein A capture step and
viral inactivation. Two polishing steps of anion and cation
… ð5Þ
exchange chromatography were then followed by ultrafiltration
and buffer exchange producing the final drug substance. The
PFD of the batch protein A platform is shown in Fig. 1. The batch
setup was modified for continuous operation with standard chro-
matography system being replaced by a Cadence™ BioSMB PD
system (Pall Corporation, USA). The protein A capture step was
ð6Þ operated in periodic countercurrent mode.30,31 The mixing tank
for viral inactivation was replaced by a CFIR and ultrafiltration–
ð7Þ diafiltration was replaced by Cadence™ inline concentrator (ILC)
and inline diafiltration (ILD) modules (Pall Corporation, USA). The
upstream process was modified to use a perfusion cell culture
process over the fed-batch process. Details of the protein A batch
and continuous processes are given in Table S4 (supporting
information).
Non-protein A platform
The non-protein A platform consisted of a upstream fed-batch
reactor followed by centrifugation as the first stage of clarification.
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Figure 2. PFDs for non-protein A platform in (a) batch mode and (b) continuous mode.
The harvest then underwent low-pH treatment for the precipita-
tion of HCP/HCD while inactivating enveloped viruses and was
depth-filtered followed by neutralization. Cation exchange chro-
matography was the capture step which also served as the pri-
mary polishing step by controlling the charge variant profile of
the mAb. This was followed by multimodal chromatography
(AEX/HIC) for further polishing. Ultrafiltration and diafiltration
were used to afford the final drug substance. The PFD for the
non-protein A platform is shown in Fig. 2. The continuous non-
protein A platform utilized a perfusion reactor to provide for con-
tinuous feed for the downstream train. Purification started with a
continuous centrifuge followed by a CFIR which was used for low-
pH treatment of the harvest. Depth filtration further clarified the
harvest and a Cadence™ BioSMB PD system (Pall Corporation,
USA) was employed for a countercurrent setup of CEX and MMC
chromatography. The final drug substance was made using
Cadence™ ILC and ILD modules (Pall Corporation, USA) to make
the process continuous. The non-protein A continuous and batch
process details are summarized in Table S5 (supporting
information).
RESULTS AND DISCUSSION
Protein A-based process platforms for mAb purification have wide
acceptance in the industry and there have been several studies
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that have examined the economic impact of continuous produc-
tion versus batch production.32,33 In this study we perform an eco-
nomic assessment of a non-protein A platform and compare it to
the traditional protein A-based continuous process platform. A
titer of 5 g L−1 and an annual scale of production of 500 kg yr−1
have been assumed.32 Optimized process parameters were used
for both platforms. As the upstream process for both the plat-
forms is the same, comparison between the downstream pro-
cesses utilized in both the platforms was the prime focus of the
economic analysis. However, while talking about COGs and also
while comparing NPV in a later section, we have included the
upstream spending to provide a holistic comparison of both the
platforms.
Protein A vs. non-protein A platform: a batch-to-batch
comparison
A novel non-protein A platform for the purification of mAbs was
recently developed in our laboratory and in this study it is com-
pared to the conventional protein A-based platform. The non-
protein A platform aims to replace the protein A capture step with
precipitation and cation exchange chromatography.
From the economic analysis data it was observed that an overall
reduction in COGs of 23% was obtained on transformation of the
batch purification process from protein A to non-protein
A. Chromatography accounted for about 80% of this reduction
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Contribution of protein A step towards cost of goods
Figure 3. Breakdown of capital and operating expenditure.
which is reflected in CapEX and OpEX comparison. There was a
34% reduction in CapEX and 45% reduction in OpEX linked with
the chromatography step. The reduction in CapEX can be attrib-
uted to the reduction in the number of chromatography steps
and hence the equipment required which is responsible for 90%
of the total reduction in CapEX. Also, as the viral inactivation step
is done in tandem with HCP/HCD precipitation during the clarifi-
cation stage, the number of tanks required in the setup is reduced
and this further contributes to the cost saving. Further breaking
up the chromatography steps, it can be seen that the polishing
chromatography contributes to 73% reduction in CapEX,
highlighting the advantage of eliminating a chromatography
step. Here it should be noted that although cation exchange chro-
matography is the capture step in the non-protein A platform, it
also simultaneously acts as the polishing step by controlling the
charge variants and reducing low-molecular-weight impurities.
Further, although the multimodal chromatography resin that fol-
lows the capture step in the non-protein A platform is more
expensive than the traditional ion exchange resins, it offers dual
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modes of interaction and thus facilitates process intensification.
Figure 4 illustrates a comparison of the unit-wise CapEX, OpEX
and COGs for protein A and non-protein A batch platforms.
Similar to CapEX, the elimination of the protein A capture step
accounted for >77% of the total reduction in OpEX. This reduction
in OpEX is equally shared by the capture and polishing chroma-
tography steps at about 40% and 38%, respectively. The major
cost savings expected from any non-protein A platform come
from elimination of protein A chromatography. The expensive
nature of protein A resin and the need for a substitute is quite evi-
dent from the high share of reduction percentage of capture chro-
matography in OpEX at a large scale. The percentage reductions
from protein A to non-protein A batch platforms are shown in
Fig. 5. The facility-dependent cost is a major share in OpEX being
64% and 77% for protein A and non-protein A platforms, respec-
tively. Elimination of the protein A chromatography step
accounted for 74% of the facility cost reduction. Even though
the non-protein A platform introduces an ultrafiltration step
before the capture chromatography, the elimination of the
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Figure 4. Unit-wise comparison of protein A and non-protein A batch platforms: (a) CapEX and OpEX comparison; (b) COGs comparison.

protein A chromatography step greatly helps reduce the facility
footprint by reducing the number of chromatography systems
required from three to two while also reducing the number of
required columns and saves on OpEX in the long run. About
70% of the total reduction in consumable cost in the non-protein
A setup is due to the replacement of protein A chromatography as
the capture step.
How beneficial is the non-protein A platform in
continuous mode?
Major economic gains were realized on transition from the con-
ventional protein A purification platform to the non-protein A
platform in the batch mode. Further, we aimed to examine if these
gains are further amplified by converting the mode of operation
from batch to continuous, as driven by the current and enormous
interest in continuous manufacturing.30,34
Figure 6 summarizes the overall benefits of continuous non-
protein A platform over batch mode. We see a 34% reduction in
COGs when the non-protein A platform is operated in a continu-
ous mode, of which 63% of the reduction is in the chromatogra-
phy step. When comparing CapEX from the non-protein A batch
to continuous setup the share of chromatography in cost reduc-
tion at large scale is about 82% which can be split into about
37% coming from the capture step and 45% coming from the pol-
ishing chromatography. Use of the novel CFIR in the second phase
6
of clarification for low-pH precipitation of HCP/HCD while simulta-
neously inactivating enveloped viruses facilitates this cost reduc-
tion. The subsequent depth filtration step resulted in improved
clarification and as a result there was a 65.7% reduction in total
OpEX. A similar trend was observed when comparing OpEX with
79% of the cost reduction attributable to chromatography split
into 36% due to capture and 43% due to polishing steps, as seen
in Fig. 7. The facility-dependent cost being the major contributor
to OpEX (about 55% of OpEX) reflected this behavior with 37%
and 45% of the facility cost reduction attributed to capture and
polishing steps, respectively. Consumable cost was the next major
share in OpEX at about 43%. There was a 23% overall reduction in
the consumable cost with the clarification and viral inactivation
steps accounting for the major share of 58% in cost reduction fol-
lowed by chromatography at 42%. The savings here were primar-
ily due to better control strategies deployed in the continuous
setup allowing the maximum use of membrane area and thus
reducing the number of filters used for clarification. The amount
of resin utilized also decreased due to the efficient use of resin
binding capacity in the continuous setup.
Is continuous non-protein A platform better than its
protein A counterpart?
We have seen the benefits of the non-protein A platform over the
protein A platform in batch mode and also the advantages of

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Contribution of protein A step towards cost of goods
Figure 5. Percentage cost reduction from protein A batch to non-protein A batch platform: (a) CapEX and OpEX; (b) facility cost; (c) consumable cost.
operating the non-protein A platform in continuous mode. How-
ever this comparison would be incomplete without talking about
the continuous protein A platform.
As the effects of converting a batch process to a continuous pro-
cess would be similar for both platforms, we expected the gains to
be less than those in batch-to-batch comparison. However, there
was a 25% reduction in COGs going from continuous protein A
platform to continuous non-protein A platform. Over 75% of the
reduction in COGs was seen to be due to chromatography steps
as seen in the unit-wise COGs comparison in Fig. 8. The share of
chromatography in CapEX reduction is about 85% and that in
OpEX reduction is about 74%. The major advantage of running a
continuous non-protein A platform is that the flow rates at which
the process operates are low enough to be accommodated by a
BioSMB pilot-scale system, whereas the continuous protein A plat-
form requires a BioSMB process-scale system which adds signifi-
cantly to CapEX. The same is reflected in OpEX and seen when
further looking at the facility-dependent cost where about 86%
of the reduction is due to the smaller BioSMB system. Figure 9
gives a comprehensive comparison of the facility and consumable
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spending for both continuous platforms. There is a 64% reduction
in consumables due to the elimination of protein A resin from the
process. Due to the simultaneous precipitation of HCP/HCD and
viral inactivation, the number of depth filters and 0.22 μm filters
required is also reduced, accounting for about 35% of consum-
able cost reduction.
Taking the leap from protein A batch platform to non-
protein A continuous platform
There are major benefits in adopting a non-protein A setup for
manufacturing of mAbs in a batch mode. And even though many
studies have compared the traditional protein A batch setup to a
protein A continuous setup, the majority of the biopharmaceuti-
cal industry still uses protein A platforms.35,36 The investments
required for converting any of the platforms to continuous mode
are the same. Both the platforms require the CFIR, BioSMB and
ILC/ILD modules. We will therefore focus on the benefits of mov-
ing from the traditional protein A batch setup to a non-protein
A continuous setup. The comparison follows the same pattern
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Figure 6. Unit-wise comparison of non-protein A batch and continuous platforms: (a) CapEX and OpEX comparison; (b) COGs comparison.
as before wherein all data are calculated based on the purification
regime while the COGs comparison includes the upstream cost.
We see the highest reduction of 39% in COGs comparing the
protein A batch platform to the non-protein A continuous plat-
form. About 70% of this reduction is due to the chromatography
step. Figures 8 and 9 give a stepwise comparison moving from
protein A batch process to protein A continuous process to non-
protein A continuous process. The share of chromatography in
CapEX reduction is about 83% and that in OpEX reduction is
79%. Even though the BioSMB PD system costs more than a stan-
dard chromatography system, the protein A batch setup requires
three systems to generate 500 kg yr−1 while the continuous non-
protein A setup can run on a single BioSMB PD system, leading to
major savings in terms of CapEX and facility-dependent cost. The
benefits of eliminating a chromatography step along with the
apparent advantage of a continuous process make the continu-
ous non-protein A platform a lucrative option.
The motive behind developing a non-protein A platform was to
reduce the costs incurred of purchasing the protein A resin. The
high cost of protein A resin translates into a heavy initial
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investment during production. A way to compare the investment
in capture chromatography media is to look at the cost of resin
per unit of product produced. As protein A resin cost is the highest
consumable cost in the purification regime, we have normalized
the cost of capture resin in each platform to the protein A resin
cost in the standard batch platform. Figure 10 shows a plot of cap-
ture resin cost per kilogram of product normalized to the protein
A resin cost for the batch platform. The normalized resin cost per
kilogram of product for protein A batch and continuous platforms
is very close after about 80 kg of product is produced. In the case
of the non-protein A platform in batch and continuous mode, the
normalized resin cost per kilogram of product overlaps after just
5 kg of production.
Although any investment will eventually pay for itself over time,
there is a very large difference in the initial investment required in
the capture resin for both the platforms. The protein A continuous
process required about 72% of the resin required for the protein A
batch setup. But the costs of capture resin for the non-protein A
batch and continuous setups are about 7% and 5% of the total
cost of protein A resin required in the batch mode. This stark
J Chem Technol Biotechnol 2021
Contribution of protein A step towards cost of goods
Figure 7. Percentage cost reduction from non-protein A batch to continuous platform: (a) CapEX and OpEX; (b) facility cost; (c) consumable cost.
difference in the initial investment is what drives research towards
non-protein A-based mAb purification platforms.
NPV analysis
NPV is an important tool for understanding the economic profitabil-
ity of any project. It provides complete understanding of economic
aspect as it takes capital investment as well as annual operational
costs into consideration. NPV considers time value of money as it is
the sum of present values of all the future cash flows in the estimated
time period of plant operations. Payback period is the number of
years required to reach the breakeven point for the initial invest-
ment. NPV analysis is a better metric than payback time because it
considers annual variation in expenses and revenue over the years.
Selling price of mAb for both the purification strategies has been
fixed at $20 mg−1. For calculating NPV, it was assumed that produc-
tion began after 3 years of capital expenditure in setting up the
plant. A discount rate of 7% per annum and income tax of 10% per
annum were used for the NPV assessment. After setup, 15 years of
plant operation has been assumed. CapEX and OpEX values were
taken from previous sections.
A shift from protein-A batch manufacturing platform to non-
protein-A batch manufacturing platform shows an advantage
(ΔNPV) of $36 million. NPV is a holistic economic evaluation of
manufacturing platforms and a gain in NPV over 15 years of oper-
ation suggests that a non-protein A-based batch facility will give
higher returns to the tune of $36 million as opposed to a protein
A-based batch facility over the same time period. Hence, NPV
gains aid decision-making for selection and development of bio-
processing technology.
In going from batch to continuous manufacturing mode for the
non-protein A purification strategy, NPV gains (ΔNPV) of $44 mil-
lion were observed. Therefore, given a choice of investing in batch
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or continuous manufacturing facilities, an investment in the con-
tinuous facility will earn $44 million extra as opposed to its batch
counterpart, over 15 years of operation.
A shift from protein A continuous manufacturing platform to
non-protein A continuous manufacturing platform shows a fur-
ther advantage (ΔNPV) of $25 million. To put this gain into per-
spective, with this advantage three new non-protein A
continuous facilities can be built, each with a throughput of
500 kg yr−1, as the CapEX requirement of an end-to-end non-
protein A continuous facility is $7.4 million (upstream and down-
stream combined). The ΔNPV values moving from one platform
to another are summarized in Table 1.
Overall, the continuous manufacturing mode of the non-protein
A purification strategy is the most cost-effective option against
protein A batch manufacturing, as it gives an advantage (ΔNPV)
of $81 million, enough to build ten new continuous non-protein
A-based mAb manufacturing facilities producing 500 kg yr−1.
The payback time for all platforms being considered is similar at
just over 3 years. However, the reason for this is the extremely
high-value product being considered.
Effect of scales on non-protein A platform: batch to
continuous
For the purpose of this study two scales were assumed at a titer of
5 g L−1. The small scale was assumed to be at 15 kg yr−1 and large
scale was assumed at 500 kg yr−1.32 Here we compare the effect
of scale on the non-protein A platform going from batch to con-
tinuous mode of operation (Fig. 11).
At the small scale we see a 32% reduction in CapEX, while at the
large scale it was 75% reduction when comparing both modes.
There was about 78% reduction in CapEX for the clarification
and viral inactivation step at small scale which can be attributed
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Figure 8. Unit-wise comparison of protein A batch, protein A continuous and non-protein A continuous platforms: (a) CapEX and OpEX comparison;
(b) COGs comparison.

to process intensification which resulted in fewer tanks and hence
a reduced facility footprint. At large scale the chromatography
step showed the highest reduction with 80% and 78% in capture
and polishing steps, respectively. This trend is somewhat reflected
when comparing Opex at both scales. There was a 33% reduction
in OpEX at small scale and about 65% reduction at large scale. The
clarification and viral inactivation step is the major contributor to
savings at 76% at small scale whereas the chromatography steps
save about 76% each at large scale.
It is common to see increased savings at clinical scales when
comparing batch and continuous manufacturing.32,37 In the case
of the non-protein A process platform, however, the reason for
the discrepancy is the chromatography equipment used at both
scales. The cost of multicolumn chromatography equipment used
at small scale increased the overall cost at that scale. However, the
same equipment can also be used for large-scale production,
unlike in batch manufacturing where larger sized equipment is
required for larger scale and costs considerably more than the
corresponding equipment used at small scale.
Sensitivity of capture step parameters
Analyzing the effect of multiple parametric uncertainties on the
10
development. Uncertainties like upstream titer, chromatographic
yield and impurity removal have been studied by other
researchers by developing an optimization algorithm to find the
best column sequencing and sizing strategy.38 Other researchers
have studied the effect of nine parameters at various levels on
specific growth rate in perfusion mode of cell culture for the pro-
duction mAbs to help in process design.39
As the aim of the present work was to emphasize the role
that protein A chromatography plays in the total cost of mAb
production, a comparative sensitivity analysis of the capture
steps was performed for both platforms. Depending on the
type of product, the interaction with the resin may change
the yield of the step and/or dynamic binding capacity (DBC)
of the resin.
The effect of capture chromatography yield was evaluated on
the overall COGs. For the protein A platform, a yield variation of
70% to 90% was considered where the optimized process yield
was 90%, whereas for the non-protein A platform a range of
75% to 95% was considered where the optimized process yield
was 95%. The analysis was performed by keeping the facility
size and other unit operation yields constant. As the yield of
the capture step decreases, the overall production rate

cost of production is of interest to those involved in process decreases resulting in an increase in the total COGs as shown

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Contribution of protein A step towards cost of goods www.soci.org

Figure 9. Unit-wise facility and consumable cost comparison of protein A batch, protein A continuous and non-protein A continuous platforms.

Figure 10. Plot of normalized capture resin cost per kilogram of product.

Resin aging is directly dependent on the DBC as the number of

Table 1. ΔNPV analysis across platforms over 15 years of operation
cycles required to process a certain amount of product increases
Non-protein A Non-protein A or decreases with the DBC in turn affecting the total amount of
batch continuous resin used. As protein A amounts to more than 70% of the total
consumable cost for the protein A platform, the effect of DBC on
Protein A batch $36 million $81 million total consumable cost was evaluated and compared with the
Protein A NA $25 million non-protein A platform. Analysis was done for both platforms by
continuous keeping the facility size constant and only changing the DBC of
Non-protein A NA $44 million the capture step. As both the capture steps operate by exploiting
batch different modes of separation, the ranges of both platforms vary
considerably. For the protein A platform in batch mode a DBC
range of 40 to 50 g L−1 was considered. For the protein A platform
in Fig. 12. The percentage increase in COGs was the same in in continuous mode, the capture step operated in periodic counter-
each range for both platforms as the overall yield also changed current mode such that the complete binding capacity can be uti-
in the same ratio. However in both batch and continuous lized by overloading the column. The unbound product was then
modes, the highest COGs achieved in the non-protein A plat- captured by the second column in series. Hence a DBC range of
form were $37 g−1 and $24 g−1 at a yield of 75% which were 64 to 80 g L−1 was considered for this mode. A DBC range of
still less than the lowest COGs in the protein A platform which 32 to 40 g L−1 was considered for the non-protein A platform in
were $41 g−1 and $27 g−1 at 90% yield.
11

both batch and continuous modes. As cation exchange

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 www.soci.org V Bansode et al.

Figure 11. Unit-wise effect of scale on non-protein A batch and continuous operation: (a) CapEX; (b) OpEX; (c) COGs.

Figure 12. Comparison of overall COGs for both platforms in batch and continuous mode at different capture step yields.

chromatography is the capture as well as polishing step in the non-
protein A platform, it cannot be operated in periodic countercur-
rent mode as it would result in loss of charge variant resolution,
and charge variant profile is a critical quality attribute for mAbs.
Hence CEX in continuous mode was loaded the same as in
batch mode.
The decrease in DBC resulted in an increase in OpEX for all plat-
forms. However, the increase was not significant, due to which the
COGs did not increase significantly. There was however a notable
12
change in the consumable cost as seen in Fig. 13. In batch mode
at a DBC of 40 g L−1, the protein A resin requirement resulted in
a 28% increase in the total consumable cost. There was no change
in protein A resin requirement at DBC values of 45 and 50 g L−1.
As the continuous mode operates by always utilizing the maxi-
mum binding capacity of the resin, the resin utilized always
increases with a decrease in DBC. The consumable cost for the
protein A platform in continuous mode increased by 7% and
16% at DBC of 72 and 64 g L−1, respectively. For the non-protein

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Contribution of protein A step towards cost of goods
Figure 13. Comparison of total consumable cost for both platforms in batch and continuous mode at different capture step DBC values.
A platform in batch mode, there was no change in the resin
requirement within the DBC range considered. In continuous
mode, the non-protein A consumable cost increased by 1% and
2% at DBC values of 36 and 32 g L−1, respectively.
Protein A resin amounts to more than 70% of the total consum-
able cost for the protein A platform. It has a lower lifetime com-
pared to CEX resins and hence has to be replaced more often.10
This is reflected in the overall consumable cost change as the resin
requirement changes.
CONCLUSIONS
The presented non-protein A continuous processing platform
provides a lucrative alternative to the standard protein A continu-
ous processing platforms. Although transforming an existing pro-
tein A batch facility to a non-protein A facility could result in extra
investment, one could consider the novel platform when building
a new facility. One could also consider investing in a new contin-
uous non-protein A facility as one of the key advantages of the
platform is that the chromatography equipment at small scale
can be employed for large-scale production thus eliminating the
need for investing in two setups. This is also evident in the NPV
comparison where the continuous non-protein A platform
showed the highest NPV gains. The non-protein A platform is flex-
ible and can be extended to a wide range of products when
coupled with single-use equipment.
ACKNOWLEDGEMENTS
The authors acknowledge funding support from the Department
of Biotechnology, Ministry of Science and Technology
(no. BT/COE/34/SP15097/2015).
CONFLICT OF INTEREST
The authors declare that they do not have any conflict of interest.
SUPPORTING INFORMATION
Supporting information may be found in the online version of this
article.
J Chem Technol Biotechnol 2021 © 2021 Society of Chemical Industry
www.soci.org
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