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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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10  
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2
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.  
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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
100  
80  
60  
40  
20  
0
25  
20  
15  
10  
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.  
80  
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
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0
10  
8
6
14  
12  
10  
8
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4
Tox DS ready  
GMP DS MFG & release  
4
2
GMP DP MFG & release  
2
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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  
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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
2
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4
2
0
2.5  
2.0  
1.5  
1.0  
0.5  
0
8
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  
8
1000  
800  
600  
400  
200  
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6
4
2
0
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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4
2
-10  
-20  
-30  
Figure 7. Empowered by our platform,  
ProBio delivers diverse CHO host cells  
to meet your needs  
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0
100  
200  
300  
400  
0
CHOK1-GenS  
CHOK1-SIA+  
clone98  
clone290  
clone335  
ProMax  
Time(h)  
ProBioCDMO.com