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\begin{document}

\title{Rapid cGMP Manufacturing of COVID-19 monoclonal antibody using stable
CHO cell pools}



\author[1]{Rita Agostinetto}%
\author[2]{Jessica Dawson}%
\author[2]{Angela Lim}%
\author[3]{Mirva Hejjaoui-simoneau}%
\author[3]{Cyril Boucher}%
\author[4]{Bernhard Valldorf}%
\author[3]{Adin Ross-gillespie}%
\author[5]{Joseph Jardine}%
\author[5]{Devin Sok}%
\author[5]{Dennis Burton}%
\author[6]{Thomas  Hassell }%
\author[7]{Hervé Broly}%
\author[3]{Wolf Palinsky}%
\author[3]{Philippe Dupraz}%
\author[6]{Mark Feinberg}%
\author[8]{Antu Dey}%
\affil[1]{Merck Serono SpA}%
\affil[2]{EMD Serono Biotech Center Inc}%
\affil[3]{Ares Trading SA}%
\affil[4]{Merck KGaA}%
\affil[5]{The Scripps Research Institute}%
\affil[6]{International Aids Vaccine Initiative}%
\affil[7]{Merck Serono SA-Corsier-sur-Vevey}%
\affil[8]{Greenlight Biosciences Inc}%


\vspace{-1em}



  
  \date{June 06, 2024}


\begingroup
\let\center\flushleft
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\selectlanguage{english}
\begin{abstract}
Therapeutic proteins, including monoclonal antibodies, are typically
manufactured using clonally-derived, stable host cell lines, since
consistent and predictable cell culture performance is highly desirable.
However, selecting and preparing banks of stable clones takes
considerable time, which inevitably extends overall development
timelines for new therapeutics by delaying the start of subsequent
activities, such as the scale-up of manufacturing processes. In the
context of the COVID-19 pandemic, with its intense pressure for
accelerated development strategies, we used a novel transposon-based
Leap-In Transposase\selectlanguage{ngerman}® system to rapidly generate high-titer stable pools
and then used them directly for large scale-manufacturing of an
anti-SARS-CoV2 monoclonal antibody under cGMP. We performed the safety
testing of our non-clonal cell bank, then used it to produce material at
a 200L-scale for pre-clinical safety studies and formulation development
work, and thereafter at 2000L scale for supply of material for a Phase 1
clinical trial. Testing demonstrated the comparability of critical
product qualities between the two scales and, more importantly, that our
final clinical trial product met all pre-set product quality
specifications. The above expediated approach provided clinically-ready
material within 4.5 months, in comparison to 12-14 months for production
of clinical trial material via the conventional approach.%
\end{abstract}\selectlanguage{ngerman}%



\sloppy


\textbf{Rapid cGMP Manufacturing of COVID-19 monoclonal antibody using
stable CHO cell pools}

\selectlanguage{ngerman}\emph{Rita Agostinetto\textsuperscript{1},Jessica
Dawson\textsuperscript{2}, Angela Lim\textsuperscript{2}, Mirva Hejjaoui
Simoneau\textsuperscript{3}, Cyril Boucher\textsuperscript{3}, Bernhard
Valldorf\textsuperscript{4}, Adin Ross-Gillespie\textsuperscript{3} ,
Joseph G. Jardine\textsuperscript{5,6,7} , Devin
Sok\textsuperscript{5,6,7}, Dennis R. Burton\textsuperscript{5,7,8},
Thomas Hassell\textsuperscript{6}, Hervé Broly\textsuperscript{3}, Wolf
Palinsky\textsuperscript{3}, Philippe Dupraz\textsuperscript{3}, Mark
Feinberg\textsuperscript{6}, Antu K. Dey\textsuperscript{6,9,} *}

\textsuperscript{1}MerckSerono S.p.A, Guidonia di Montecello, Italy

\textsuperscript{2}EMD Serono, Billerica, MA, USA

\textsuperscript{3}Ares Trading SA/Merck SA Switzerland, Aubonne
Switzerland

\textsuperscript{4}Merck KGaA, Darmstadt, Germany

\textsuperscript{5}Department of Immunology and Microbiology, The
Scripps Research Institute, La Jolla, CA, USA

\textsuperscript{6}IAVI, 125 Broad Street, New York, NY, USA

\textsuperscript{7}IAVI, Neutralizing Antibody Center, The Scripps
Research Institute, La Jolla, CA, USA

\textsuperscript{8}Ragon Institute of MGH, MIT and Harvard, Cambridge,
MA, USA

\textsuperscript{9}Current address: GreenLight Biosciences Inc., 200
Boston Avenue, Suite 1000, Medford, MA, USA.

*Correspondence: Antu K. Dey; Email: adey@greenlightbio.com



\textbf{KEYWORDS}

cGMP manufacturing, CHO pools, COVID-19, monoclonal antibody

\textbf{ABSTRACT}

Therapeutic proteins, including monoclonal antibodies, are typically
manufactured using clonally-derived, stable host cell lines, since
consistent and predictable cell culture performance is highly desirable.
However, selecting and preparing banks of stable clones takes
considerable time, which inevitably extends overall development
timelines for new therapeutics by delaying the start of subsequent
activities, such as the scale-up of manufacturing processes. In the
context of the COVID-19 pandemic, with its intense pressure for
accelerated development strategies, we used a novel transposon-based
Leap-In Transposase\textsuperscript{®} system to rapidly generate
high-titer stable pools and then used them directly for large
scale-manufacturing of an anti-SARS-CoV2 monoclonal antibody under cGMP.
We performed the safety testing of our non-clonal cell bank, then used
it to produce material at a 200L-scale for pre-clinical safety studies
and formulation development work, and thereafter at 2000L scale for
supply of material for a Phase 1 clinical trial. Testing demonstrated
the comparability of critical product qualities between the two scales
and, more importantly, that our final clinical trial product met all
pre-set product quality specifications. The above expediated approach
provided clinically-ready material within 4.5 months, in comparison to
12-14 months for production of clinical trial material via the
conventional approach.



\textbf{COMMUNICATION TO THE EDITOR}

Multiple cell line sources are used as hosts to produce recombinant
prophylactic and therapeutic proteins for human use. Recombinant CHO
cell lines (Puck et al., 1958) remain a preferred host due to the
reliability, robustness and maturity of the technology in generating
clonally-derived cell line. Despite the high level of production,
batch-to-batch consistency and robustness of a clonally-derived CHO cell
line, the approach is challenging due to the burden of time and
resources required for stable clone isolation, selection, and provision
of tested cell banks for cGMP manufacturing. To overcome this challenge,
we wanted to evaluate the possibility of using non-clonal stable CHO
cell pools to expediate production of a monoclonal antibody (mAb)
against SARS-CoV2, CC6.35, particularly through cGMP manufacturing of a
single batch of material for a Phase 1 clinical trial. Since this
approach has not been rigorously tested for scaled-up cGMP manufacturing
and concerns remain that the cellular and genetic heterogeneity of
non-clonal stable CHO cell pools may result in production variability
and concomitant heterogeneous product qualities between batches, we
present a case study wherein we used non-clonal qualified cell banks and
platform processes to accelerate manufacturing of CC6.35 mAb. To do so,
we used the novel transposon-based Leap-In
Transposase\textsuperscript{®} system (Rajendra et al., 2017; Rajendran
et al., 2021) for the development of stable CHO cell lines. The
codon-optimized DNA sequence encoding the amino acid sequence for the
Heavy chain(HC) and Light chain(LC) of CC6.35 mAb along with
corresponding signal peptide and the novel expression constructs based
on the Leap-In transposon® system were designed and synthesized. These
synthesized DNA constructs along with transposase mRNA (Rajendran et
al., 2021; Wilson et al., 2007) were used to co-transfect HD-BIOP3
glutamine synthase (GS) knock-out CHO-K1 host cells. Two promoter
components were used to generate two unique sets of CHO cell pools: one
using the EF1 promoter and another using the CMV promoter.
Post-transfection the recovery of these two CHO cell pools was
performed; after the initial recovery phase, the positive CHO cell pools
were selected by outgrowth in a glutamine-free formulation at 37°C in
5\% CO\textsubscript{2} and 70-80\% relative humidity and expanded
further before cryopreservation and generation of Research Cell Banks
(RCBs). In order to estimate productivity of the CC6.35
antibody-expressing stable CHO cell pools, both cell pools were expanded
in a small-scale cell culture based on fed-batch process. After 14-days,
the harvested supernatant was measured for titer by binding to a
Protein-A biosensor on the Octet\textsuperscript{®} System.\textbf{Table
1} shows the percent (\%) viability and expression levels (in g/L) for
both EF1-CC6.35 and CMV-CC6.35 stable CHO cell pools.

Based on the higher titer, the CMV-CC6.35 cryopreserved RCB vials were
advanced for use in scale-up production. This (CMV-CC6.35) RCB was
tested for safety and regulatory acceptance for producing the two
pivotal lots of material: one for preclinical safety studies and another
for Phase 1 clinical trial.

For producing material for pre-clinical safety study, the RCB vials were
thawed and expanded using a seed-train to support a 200~L bioreactor.
The key cell culture process indicators included cell growth, cell
viability, metabolic profiles (for ammonia and lactate), bioreactor
regulation profiles (for pH, glucose, osmolality and
pCO\textsubscript{2}) and antibody titer. After 14 days, the cell
culture supernatant was harvested, clarified, and subjected to a
3-column chromatography platform purification process (involving
Protein-A column chromatography, Anion-Exchange Column Chromatography
and a Cation-Exchange column chromatography) steps to generate purified
Drug Substance (DS), formulated at 20~mg/mL in an antibody platform
buffer as Drug Product (DP) for intra-venous administration.

For producing material for the Phase 1 clinical study, a similar
upstream (cell culture) process strategy was employed except that in
this case, the RCB vials were thawed and expanded using a seed-train to
support a 2000~L bioreactor. After 14 days, the supernatant from the
2000~L bioreactor was harvested, clarified, and purified via the same
3-column chromatography platform process to generate purified Drug
Substance (DS), formulated at 20~mg/mL in a histidine-based buffer as
Drug Product (DP) for intra-venous (i.v.) administration.

Both Drug Substances and Drug Products (200L and 2000 L scales) were
tested based on a proposed analytical panel and results compared to
agreed acceptance criteria. The analytical panel, comprising of
quantitative test (protein concentration and biological activity),
qualitative tests (physico-chemical and microbiological purity) and
compendial methods, was set based on prior knowledge on antibodies of
similar structure, and after confirmation of their suitability for the
purpose. In addition, a formal analytical comparability was performed to
ensure that the two products, from 200L and 2000L scales, are
''essentially similar'' despite the changes in their production scale
and minor change in processes. The results of the analytical
comparability study are reported in \textbf{Table 2} .

In addition to results in \textbf{Table 2} , Circular Dichroism (CD)
analysis showed that both DS materials are comparable in terms of
secondary and tertiary structures (data not shown). Thermal analysis (by
Differential Scanning Fluorimetry) showed that both DS samples have
similar thermal denaturation profiles with a temperature of onset
(T\textsubscript{onset}) at 60-61~°C with two inflection points (IP) for
both samples, one at 67~°C and another at 78~°C (data not shown).
Extensive characterization of glycans (glycan mapping) of the two DS
materials showed that the various detected N-linked glycan species are
comparable, except for some minor differences in relative distribution
of the galactosylated, fucosylated and sialylated species (data not
shown). Additionally, LC-MS analysis was performed on the two DS
materials for intact mass and deglycosylated/reduced mass. Intact mass
LC-MS analysis showed that the main proteoform for both non-cGMP DS
batch and the cGMP DS materials was the intact molecule PyroQ-LC +
HC{[}-K{]} coupled with FA2-FA2 glycans (data not shown). With respect
to the expected mass of 150622.60 Dalton, the non-cGMP DS had an
experimental mass of 150622.35 Dalton, whereas the cGMP DS had a mass of
150623.80 Dalton. Some other fragments were detected and identified in
both samples, amongst which the most abundant was the LC-LC dimer (data
not shown). After deglycosylation/reduction, the LC-MS analysis revealed
that the two DS materials were overall comparable in terms of intact
molecule, except for slightly higher levels of HC C-terminal truncation
in the cGMP batch (data not shown). Finally, non-reducing peptide
mapping by LC-MS/MS showed that the (nine) detected disulphide bridges
were aligned in line with expected (canonical) ones.

In summary, we demonstrate that using non-clonal stable CHO cell pools
and use of platform processes can expediate early clinical development
of monoclonal antibodies during pandemic outbreaks of emerging
infectious such as COVID-19. We show that the antibodies produced from
these stable CHO pools at two large scales bioreactors were comparable
using platform upstream and downstream processes, and through robust
analytical tested they were deemed suitable for clinical use. Since
clinical development timeline continue to thwart rapid evaluation of
therapeutic and prophylactic interventions during pandemics and
improvements in development span essential (Kelley, 2020), we believe
that this approach of using non-clonal stable CHO cell pools, which
enabled manufacturing of early clinical trial material within 4.5
months, is a feasible alternative for rapid cGMP manufacturing, and a
means that can accelerate the pace of therapeutic and prophylactic
protein evaluation in clinic.



\textbf{ACKNOWLEDGEMENT}

We thank the various members of the Project Team at Merck KGaA and IAVI
for their contributions. We thank Sowmya Rajendran, Lynn Webster, Surya
Karunakaran and Ferenc Boldog at ATUM for Cell Line Development of the
stable CHO cell line pools. We thank the members of the senior
management team at Merck KGaA and IAVI for their support and guidance
through the project. The authors have no conflict of interest to
declare.



\textbf{REFERENCES}

Kelley, B. 2020. Developing therapeutic monoclonal antibodies at
pandemic pace. \emph{Nat Biotechnol} 38:540-545.

Puck, T.T., S.J. Cieciura, and A. Robinson. 1958. Genetics of somatic
mammalian cells. III. Long-term cultivation of euploid cells from human
and animal subjects. \emph{J Exp Med} 108:945-956.

Rajendra, Y., S. Balasubramanian, R.B. Peery, J.R. Swartling, N.A.
McCracken, D.L. Norris, C.C. Frye, and G.C. Barnard. 2017. Bioreactor
scale up and protein product quality characterization of piggyBac
transposon derived CHO pools. \emph{Biotechnol Prog} 33:534-540.

Rajendran, S., S. Balasubramanian, L. Webster, M. Lee, D. Vavilala, N.
Kulikov, J. Choi, C. Tang, M. Hunter, R. Wang, H. Kaur, S. Karunakaran,
V. Sitaraman, J. Minshull, and F. Boldog. 2021. Accelerating and
de-risking CMC development with transposon-derived manufacturing cell
lines. \emph{Biotechnol Bioeng}

Wilson, M.H., C.J. Coates, and A.L. George, Jr. 2007. PiggyBac
transposon-mediated gene transfer in human cells. \emph{Mol
Ther}15:139-145.

\textbf{TABLES}

\textbf{Table 1.} \textbf{Viability and expression (at day 14) of
non-clonal stable CHO cell pools expressing CC6.35 mAb in a fed-batch
culture.}\selectlanguage{english}
\begin{longtable}[]{@{}lll@{}}
\toprule
\textbf{CHO Cell Pools} & \textbf{Viability (in \%)} &
\textbf{Expression level (in g/L)}\tabularnewline
\midrule
\endhead
EF1-CC6.35 & 99.44 & 3.05\tabularnewline
CMV-CC6.35 & 99.20 & 6.01\tabularnewline
\bottomrule
\end{longtable}

\textbf{Table 2. Comparison of parameters of the Drug Substances from
200L (non-cGMP) and 2000L (cGMP) production runs.}\selectlanguage{english}
\begin{longtable}[]{@{}llll@{}}
\toprule
\begin{minipage}[b]{0.22\columnwidth}\raggedright\strut
\textbf{Parameters}\strut
\end{minipage} & \begin{minipage}[b]{0.22\columnwidth}\raggedright\strut
\textbf{Specifications}\strut
\end{minipage} & \begin{minipage}[b]{0.22\columnwidth}\raggedright\strut
\textbf{DS (from 200L)}\strut
\end{minipage} & \begin{minipage}[b]{0.22\columnwidth}\raggedright\strut
\textbf{DS (from 2000L)}\strut
\end{minipage}\tabularnewline
\midrule
\endhead
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Appearance\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Practically free from visible particles\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Practically free from visible particles\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Practically free from visible particles\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Clarity and Degree of Opalescence (NTU)\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 18 NTU\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
5\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
3.4\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Degree of coloration\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Not more intensely colored than Reference Solution Y5\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{}Y6\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{}Y5\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
pH\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
5.0 -6.0\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
5.6\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
5.5\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Osmolality\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
250 -400 mOsmol/Kg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
310\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
320\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Protein concentration\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
18-22mg/mL\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
20.6 mg/mL\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
19.8 mg/mL\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Biological activity (ELISA)\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
70-130 \% of Ref. STD\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
95\%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
105\%\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Purity (Reduced CGE-SDS)\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NLT 85 \%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
95.3\%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
96.2\%\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
LMW (Non-Reduced CGE-SDS)\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 12 \%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
8.3\%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
7.5\%\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.24\columnwidth}\raggedright\strut
\% Purity by iCE\strut
\end{minipage} & \begin{minipage}[t]{0.24\columnwidth}\raggedright\strut
CPI Cluster 3 (Main Peak): 97-103 \% Acidic cluster: Report Result Main
Peak: Report Result Basic Cluster: Report Result\strut
\end{minipage} & \begin{minipage}[t]{0.24\columnwidth}\raggedright\strut
CPI\%=100 Acidic Cluster: 51.0\% Main Peak: 39.1 \% Basic Cluster:
9.9\%\strut
\end{minipage} & \begin{minipage}[t]{0.24\columnwidth}\raggedright\strut
CPI\%=100 Acidic Cluster: 49.4\% Main Peak: 40.5 \% Basic Cluster:
10.1\%\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
HMW species (SE-HPLC)\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 6.5 \%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
4.3\%\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
3.8\%\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Residual HCP\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 30 ng/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
1.794 ng/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{} 0.303 ng/mg\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Residual DNA\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 5 pg/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{}0.421 pg/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{}0.22 pg/mg\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Residual Protein A\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 30 ng/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{} 0.311 ng/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{} 0.8 ng/mg\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Bioburden\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 1 CFU/10mL\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
0 CFU/ 10 mL\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
0 CFU/ 10 mL\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
Bacterial Endotoxin Test (BET)\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
NMT 0.1 EU/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{}0,01 EU/mg\strut
\end{minipage} & \begin{minipage}[t]{0.22\columnwidth}\raggedright\strut
\textless{}0,01 EU/mg\strut
\end{minipage}\tabularnewline
\bottomrule
\end{longtable}

\textbf{Hosted file}

\verb`Table1_Rita Agostinetto et al.docx`  available at \url{https://authorea.com/users/435412/articles/538453-rapid-cgmp-manufacturing-of-covid-19-monoclonal-antibody-using-stable-cho-cell-pools}

\textbf{Hosted file}

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