Premium-quality mAbs ranging from 1-100 g in 3-4 weeks
Large-Scale Monoclonal Antibody Production GramExpress Platform: Qian Guo, Ning Xu, Li Zhang, Jiansheng Wu WuXi Biologics, NO.240 Hedan Road, Pudong New District, Shanghai, China (Contact: PS_Marketing@wuxibiologics.com)
Abstract
Methods All experiments were conducted within WuXi Biologics' Protein Sciences (PS) Department. The DNA sequence codon underwent optimization using WuXi Biologics’ laboratory data management system (LDMS), and the plasmid was then constructed into our proprietary vector. Subsequently, a WuXian TM Transient procedure was performed in adherence to SOPs, and the culture medium was collected a fter 7 days. A fter clarifying the culture medium, all supernatant was loaded onto suitable purification columns based on the titer. Purification steps involved ProA a ffinity capture, size exclusion chromatography (SEC), and ion exchange (IEX). The final product underwent analysis using SDS-PAGE, SEC-HPLC, intact Mass analysis, and endotoxin detection.
Monoclonal antibodies (mAbs) continue to be a major focus in the pharmaceutical industry, with their development advancing at a fast pace. Meeting customer demand by e fficiently producing premium-quality antibodies on a large scale in a short timeframe presents a significant challenge. At WuXi Biologics, our Protein Sciences (PS) Department has developed the GramExpress platform to meet this challenge. By harnessing our high-titer WuXian TM Transient expression system and adaptable purification methods, GramExpress achieves superior yields and purity within an accelerated timeframe. Introduction Typically, producing gram-level mAbs involves stable pool production, a process that takes at least 4 weeks for expression. To expedite this process, the PS Department dedicated several years to innovating and enhancing transient expression. Our aim was to achieve higher titers in less time. By optimizing codons, host cells, and the cell culture process, we significantly improved the titer of our expression system, increasing it from 0.15 g/L to 2.2 g/L (based on an internal control mAb). Furthermore, we shortened the timeline from 10 days to 7 days. In the purification stage, we employ 2-3 steps involving columns of di ffe rent sizes to obtain the desired high-quality product. Our GramExpress platform is capable of producing 1-100 g of high quality mAbs within a 3-4 week timeframe. WuXian TM Transient
Results
Figure 4: An Example of MAB Purified Using 3-Column Steps at 60 L Scale & Completed within 4 Weeks
Key Features of the GramExpress Platform
• Ultra-high titer
WuXian
TM Transient
expression
• Delivers 1-100 g mAbs in 3-4 weeks • Premium-quality (purity >98%, endotoxin <0.2 EU/mg)
Within a span of 4 weeks (1 week for DNA preparation, 1 week for expression, and 2 weeks for purification and QC), ~79 grams of target protein was produced, exhibiting a purity of 99% via SEC-HPLC, an endotoxin level of <0.02 EU/mg, and an overall recovery rate of 76%.
The Development of the WuXian Transient TM System Figure 1: WuXian Transient TM Development Timeline
Figure 5: An Example of Antibody Purified Using 2-Column Steps at 10 L Scale & Completed within 3 Weeks
2024 WT 3.0
2022
2014
WT 3.0
WT 4.0
WT 2.5
WT 1.0
WT 2.0
Control mAb Titer
Ongoing
2.6 g/L 7 Days
2.2 g/L 7 Days
0.15 g/L 10 Days
0.6 g/L 7 Days
1 g/L 7 Days
Figure 2: Optimization Strategies for WuXian Transient TM WuXian Transient TM : Strategies for Optimization
Expression to Support
High-Titer Expression
~12 grams of pure mAb was produced within a 3-week timeframe (1 week for DNA preparation, 1 week for expression, and 1 week for purification and QC), exhibiting a 99% purity via SEC-HPLC and an endotoxin level of <0.16 EU/mg.
Conclusions
WuXi Biologics' Protein Sciences Department has devised strategies for gram-level mAb production that ensures increased yield, enhanced purity, and an even shorter production timeline.
The optimization process encompassed multiple stages, commencing with bioinformatics and host clone selection, and extending through transfection and cell culture phases, all aimed at achieving ultra-high titer expression.
WuXian Transient TM : Strategies for Optimization
We thank all scientists from Department of Protein Sciences of WuXi Biologics for their great support during the method development. We also thank legal department, public relationship department and marketing department for their great help in patent filing and poster preparation and revision. Acknowledgments
Figure 3: High Titer of Control Antibody Expression
A commercial antibody was produced using WuXian Transient TM 3.0 system with multiple batches as a control, demonstrating high titer expression on both Day 4 and Day 7 of culture.
Control Antibody
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About WuXi Biologics CRO Services
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WuXi Biologics is a leading global CRDMO, with the R represented by our premium-quality CRO Services. With expertise in diverse drug modalities, we deliver comprehensive research services, including anti- body discovery, protein production and engineering, and in vitro/in vivo testing, to seamlessly advance our clients’ molecules from concept to CMC development. Drug Development Expertise Empowering Research Services for Biologics
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For more information see us at https://www.wuxibiologics.com/cro-services
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GramExpress_Poster_202509V1
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To discuss this poster: PS_Marketing@wuxibiologics.com
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