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_China@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 after 7 days. After clarifying the culture medium, all supernatant was loaded onto suitable purification columns based on the titer. Purification steps involved ProA affinity 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 efficiently 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 WuXian TM Transient 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 different 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.
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 System Figure 1: WuXian TM Transient Development Timeline
Figure 5: An Example of Antibody Purified Using 2-Column Steps at 10 L Scale & Completed within 3 Weeks
Expression Titer Culture Days
WuXian Transient: Strategies for Optimization Figure 2: Optimization Strategies for WuXian TM Transient 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: Strategies for Optimization Figure 3: High Titer of Control Antibody Expression
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 WuXi Biologics is a global leading Contract Research, Development and Manufacturing Organization (CRDMO) offering end-to-end solutions to enable partners to discover, develop and manufacture biologics from concept to commercialization. About WuXi Biologics For more information see us at wuxibiologics.com
A commercial antibody was produced using WuXian Transient 3.0 system with multiple batches as a control, demonstrating high titer expression on both Day 4 and Day 7 of culture.
Poster modified on 1/9/2024
To discuss this poster: PS_BD@wuxibiologics.com
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