Poster
Integrated DNA-to-IND Workflow Overview: ProMax and ExpressCMC
TM
Enabling ExpressCMC with Next Generation Cell
Line Platform
Ying Li, Yeqiong Li, Lei Zhang, Yue Sheng, Felix Zhu
ProBio, No. 28 Yongxi Road, Jiangning District, Nanjing, Jiangsu, China
48h Post-transfection
Cell pool fed-batch
clone fed-batch
INTRODUCTION
Cell pool fed-batch
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The development of stable, high-producing mammalian cell lines remains a pivotal challenge in
bioprocessing. Traditional approaches often force a difficult trade-off: achieving high initial product
titers can come at the expense of long-term genetic stability, leading to unpredictable productivity
drops during extended culture and scale-up. This instability injects significant uncertainty and risk
into downstream process development and manufacturing. It necessitates lengthy clone screening
campaigns, extensive stability studies, and complex mitigation strategies, ultimately prolonging
development timelines and consuming valuable resources.
0
0
Transfection1
Transfection2
Top 8 pools
Top 12 clones
SEC-HPLC
CE-SDS-NR
Cell pool
Clones
Peak VCD
100
80
60
40
20
0
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80
60
40
20
0
25
20
15
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5
Cell pool
Clones
clone fed-batch
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Here, ProBio introduces the ProMax Next-Generation Expression System, a novel solution engineered
to break the traditional trade-off between high titer and long-term stability.
HMW
Main
LMW
Other
Main
Cell pool
Clones
icIEF
CE-SDS-R
Harvest VIA
80
60
40
20
0
100
100
80
60
40
20
0
Cell pool
Clones
Cell pool
Clones
Leveraging this robust platform, we have launched our ExpressCMC™ service. The predictable,
high-yield, and stable cell clones generated by the ProMax platform enable an accelerated,
de-risked development path. We demonstrate rapid cell pool recovery, consistent critical quality
attributes (CQAs) between pool and clonal derivatives, and a streamlined timeline to deliver
Toxicity Batch material in 4 months and GMP Drug Product in 9 months. This integrated solution
offers a new paradigm in bioprocessing efficiency, reducing time to clinic while ensuring scalable,
high-quality production.
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60
40
20
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Acidic
Main
Basic
HC
LC
LC+HC Other
Cell pool
Clones
Figure 5. Consistent critical attributes between ProMax-derived pools and clones
Data demonstrate the high consistency in both process performance and product quality
between production-ready cell pools and single-cell-derived clones generated using the ProMax
platform. This consistency, achieved without extensive screening, validates the platform's ability
to rapidly deliver clonal cell lines that faithfully represent the performance profile of the
early-stage pool, derisking scale-up and tech transfer.
PROMAX VECTOR
Culture
Harvest VIA Harvest Lac
Peak VCD
Temp.Shift
Timing
Cedex IgG
Duration
Molecular
Vector
Temp.
(106 cells/mL)
(%)
(g/L)
(g/L)
(day)
TiterLift™ titer boosting element
mAb1
mAb1
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
ProMax
36.5 33
36.5 33
36.5 33
36.5 35
36.5 35
36.5 35
36.5 33
36.5 33
36.5 33
36.5 35
36.5 35
36.5 35
D5
D6
D7
D5
D6
D7
D5
D6
D7
D5
D6
D7
14
14
14
14
14
14
14
14
14
14
14
14
18.6
18.0
18.6
21.5
21.9
83.85
84.67
84.20
85.58
84.36
79.99
94.95
92.10
91.95
0.09
0.10
0.09
0.10
0.11
5.32
5.14
Significantly High Titer
The TiterLift™ element enables superior
mAb1
mAb1
mAb1
5.48
5.61
integration efficiency, rapidly generating
clones with high transgene copy numbers,
5.49
5.45
6.44
6.48
6.25
6.22
6.29
6.43
Ideal for All Molecules
which directly boost titer
mAb1
19.9
0.12
0.09
0.09
0.09
0.13
0.14
0.14
Achieve consistently high
bsAb2
bsAb2
bsAb2
bsAb2
bsAb2
bsAb2
25.5
30.0
28.3
27.6
27.4
27.8
ProMax
expression across
SteadyHold™ stability enhancing
generations, regardless of
element
molecule type.
92.18
90.18
89.88
Exceptional Long-Term Stability
The SteadyHold™ element sustains
transgene expression activity, preventing
the transgene from being switched off
over time, resulting in minimal titer drop
(<30%) over 60+ generations without
selective pressure.
Table 1. Process robustness of ProMax
Neither the temp. shift timing nor temp. shift setpoint showed a significant impact on VCD, VIA,
lactate or titer. Lactate accumulation was not observed for all conditions. ProMax's robustness
bypasses traditional process optimization, reducing process development timelines.
Figure 1. The schematic of ProMax ve
The TiterLift™ element drives superior integd isolation of high-producing
clones with elevated transgene copy numy-leading initial titers. The
SteadyHold™ element maintains transcriptionsgene silencing to ensure
exceptional production stability, providing a stacturing. Protected Innovation:
This technology is the subject of a pending U.S. patent application (No. 63/652,263), underscoring its
novel design and commercial value.
EXPRESSCMC™
Timeline (Month)
1
2
3
4
5
6
7
8
9
Cell line development*
RESULTS
Process development
Formulation development
Clone fed-batch of Protein
Clone fed-batch of MAB
Clone fed-batch of BSAB
Average Titer
Quality study & method qualification
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4
2
0
10
8
6
14
12
10
8
6
4
Tox DS ready
GMP DS MFG & release
4
2
GMP DP MFG & release
2
0
0
* From DNA synthesis to Top 1 clone
Protein 1
Protein 2
MAB 1
MAB 2
BASB 1
BASB 2
mAb
BsAb
Protein
*The titers are not process optimized
Applicable to
Get Tox Batch Ready in 4 Months and GMP DP in 9 Months
• mAb and symmetric bsAb
Figure 2. The ProMax platform enables high-titer production across multiple modalities
• Liquid injection
• DP concentration ≤ 50mg/ml (mAb)
Bar graphs demonstrate the robust protein expression achieved by representative clones generated
with the ProMax expression system. The platform consistently delivers high titers for multiple
therapeutic modalities, with peak performance exceeding industry benchmarks. mAb titer: average
7.53 g/L, up to 15.23 g/L, bsAb titer: average 6.97 g/L, up to 10 g/L, recombinant protein titer: average
5.8 g/L, up to 14.54 g/L.
Empowered by proprietary ProMax vector
• DP concentration ≤ 30mg/ml (bsAb)
Figure 6. Express CMC™: industry-leading development speed
Gantt chart illustrates the streamlined, de-risked development and manufacturing pathway
enabled by the predictable performance of the ProMax expression system. By eliminating the
traditional bottlenecks and variability associated with unstable cell lines, we offer a de-risked,
accelerated path from DNA to clinic. Our proven timeline—delivering Toxicity Batches in 4
months and GMP Drug Product in 9 months—enables partners to compress development
schedules, reduce resource burdens, and accelerate candidate progression with greater
confidence.
Pool recovery time of Protein
Pool recovery time of MAB
Pool recovery time of BSAB
25
20
15
10
5
20
15
10
5
15
10
5
Protein 1
Protein 2
MAB 1
MAB 2
BSAB 1
BSAB 2
0
0
0
Ref.
ProMax
Ref.
ProMax
Ref.
ProMax
Ref.
ProMax
Ref.
ProMax
Ref.
ProMax
CHO HOST CELL OPTIONS
Case study of CHOK1-ADCC+ cell line
Figure 3. Rapid generation of cell pools with the ProMax platform
The TiterLift™ element drives superior genomic integration efficiency in transfected cells, while the
SteadyHold™ element sustains transgene expression activity. This synergistic action enables the rapid
induction of stable, high-producing pools, forming a critical foundation for accelerated development
workflows.
CD16a 158V/F affinity (BLI)
PBMC-based ADCC
CHO-K1 Parental Cells
MAB1 clone stability
MAB2 clone stability
Protein1 clone stability
Protein2 clone stability
Cell pool
Clones
PDL0
PDL0
PDL0
PDL60
PDL0
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6
4
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6
4
2
0
2.5
2.0
1.5
1.0
0.5
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6
4
2
0
PDL60
PDL0
PDL60
PDL60
PDL0
PDL60
PDL0
PDL60
PDL60
PDL60
PDL60
PDL0
1. CHOK1-GenS: Wild type cell line from ECACC
2.CHOK1-ADCC+: Proprietary cell line system to
produce afucosylated antibodies
PDL0
PDL0
PDL60
PDL60
PDL60
PDL0
PDL0
Case study of CHOK1-SIA+ cell line
3.CHOK1-SIA+：Independently developed cell line
engineered for superior serum half-life and
pharmacokinetics through elevated, human-like
terminal sialylation.
Protein titer
Protein sialylation
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1000
800
600
400
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clone84
clone95
clone311
clone30
clone183
clone223
clone75
clone79
clone125
clone75
clone103
clone134
• Suspension-adapted CHO-K1
Figure 4. Exceptional long-term stability of
ProMax-derived clones
• Animal product-free culturing
CHOK1-GenS
CHOK1-SIA+
CHOK1-GenS
CHOK1-SIA+
• Compatible with chemically defined com-
mercial media
BSAB2 clone stability
clone stability
ProMax demonstrated stability in all tested clones, with a
titer drop of <30% in stability studies over 60+
generations without selective pressure. This predictable
and minimal productivity decline de-risks manufacturing
campaigns, ensures reliable product supply, and is a
fundamental enabler of the accelerated ExpressCMC™
timeline.
Rat PK
Rat PK
10
8
6
30
20
10
PDL0
PDL0PDL60
1000
100
10
250
200
150
100
50
PDL0
CHOK1-GenS DS,20 mg/kg
CHOK1-SIA+ DS,20 mg/kg
PDL60
PDL60
• Versatility for biopharmaceutical applications :
potential for customization
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2
-10
-20
-30
Figure 7. Empowered by our platform,
ProBio delivers diverse CHO host cells
to meet your needs
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100
200
300
400
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CHOK1-GenS
CHOK1-SIA+
clone98
clone290
clone335
ProMax
Time(h)
ProBioCDMO.com