Low aggregates (>95% monomer) and low endotoxin (<0.5EU/mg) antibody production in just 3 weeks at the scale of 1-100mg, suitable for both in vitro and in vivo studies.
Accelerating Early Drug Discovery Process with Vanilla Antibody Production using 777 Platform
Gaozhan Zhang, Deyang Zeng, Dawei Zhang, Mengjie Lu, Jiansheng Wu WuXi Biologics, NO.240 Hedan Road, Pudong New District, Shanghai, China (Contact: PS_BD@wuxibiologics.com)
There are thousands of candidates in early antibody drug discovery process. The assessment of binding affinity, biological properties, and critical developability parameters often requires small amounts of purified materials (<1 g). However, the lack of high-throughput (HTP) methods for the generation of vanilla antibodies has resulted in a bottleneck that has hampered their therapeutic evaluation. To support our clients and overcome the challenges associates with protein production, WuXi Biologics’ Protein Sciences Department has established the HTP antibody production platform: 777. The name 777 represents 7 days for gene synthesis, 7 days for protein expression, and 7 days for purification and QC. 777 is an industry-leading platform that can deliver vanilla antibodies with high quality (>95%), low endotoxin (<0.5 EU/mg) within 3 weeks. The successful establishment of our 777 platform greatly shortens the timeline of HTP production of vanilla antibodies for the screening of best candidates, accelerating the process of early-stage drug discovery. Abstract
Case study: SEC-HPLC and Caliper-SDS data In this case study, the purity of vanilla antibodies, which were produced with the 777 platform, showed high purity.
Fig 5. SEC-HPLC and Caliper-SDS data of vanilla antibodies produced in the 777 platform
Introduction
Vanilla antibody represents a cluster of immunoglobulin proteins that can bind to protein A and/or protein G. In the early stage, it often requires the screening of >1000 vanilla antibodies for the assessment of binding affinity, biological properties, and critical developability parameters. While typically the timeline for vanilla protein production is about 7-8 weeks. Shortening the timeline and increasing the throughput are the main challenges in early-stage drug discovery.
Fig 1. The challenges in vanilla antibody production
(A) Vanilla Antibody • huIgG1, huIgG4
(C)
• mIgG1, mIgG2a, mIgG2b • Rabbit IgG • Rat IgG2a, rat IgG2b
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Protein Expression
Gene Synthesis
7-8 Weeks
Protein Purification
Protein Analytics
Case study: 72 Regular IgG at 20 mL – 4-Day Transfection Sometimes our clients don’t need much protein as well as high purity. Our 777 platform can adjust the process and meet client’s requirements. In this case study, the duration of WuXian Transient was shortened to 4 days. Furthermore, only one-step purification was performed.
(A) The classification of vanilla antibody. (B) The typical timeline for vanilla antibody production. (C) Developability for early discovery. There are thousands of candidates that need to be produced for therapeutic evaluation.
Fig 6. The scope of the 777 platform can be customized
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Fig 2. The scheme of the 777 platform
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7 days for Gene Synthesis 7 days for Protein Expression 7 days for Purification and QC
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With the empowerment of Protein Sciences’ automation technology, the 777 platform can deliver vanilla antibodies with quality within 3 weeks.
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The protein amount and quality were slightly decreased when the duration and purification steps were changed to 4 days and one-step respectively.
Fig 3. The features of the 777 platform
Gene synthesis, 7 days
Protein Expression, 7 days
Purification and QC, 7 days
• WuXi Biologics’ Protein Sciences Department has developed a high-throughput vanilla antibody production platform 777 which can support early drug-discovery projects. • The 777 platform can deliver small amounts of high-quality vanilla antibodies which are suitable for in vivo and in vitro studies. • Thousands of vanilla antibodies have been produced utilizing our 777 platform and data shows the platform is robustness, time-saving and reproducible. Conclusion All experiments were conducted within WuXi Biologics’ Protein Sciences Department. Briefly, the codon of DNA sequence was optimized by WuXi Biologics lab data management system (LDMS) and the plasmid was constructed into WuXi Biologics proprietary vector. WuXian Transient was performed based on SOP and the culture medium was collected after 7 days (4 days in some cases). After centrifugation, all supernatant was loaded onto an appropriate tandem protein A and SEC column. The final product was analyzed by Caliper-SDS, SEC-HPLC and endotoxin detection. Method
• WuXi Bio Codon • HTP automation platform for gene synthesis
• WuXian TM Transient • Multi-scales: 20 mL- 500 mL • High titer: 3 mg - 1 g
• Automated 2-step purification (AC-SEC) • Low aggregation (>95% monomer) • Low endotoxin (<0.5 EU/mg)
With high capacity of gene synthesis, multi-scale of WuXian Transient and automated 2-step purification, the 777 platform can produce high quality vanilla antibodies which are good for in vivo and in vitro studies.
Results
Case study: 170 vanilla IgG at 20 mL In this case study, 170 vanilla antibodies were produced with the 777 platform. Each antibody was expressed at 20 mL scale, and the average titer was 0.7 g/L. After 2-step automated purification, enough proteins (average = 4.5 mg) were obtained with <0.2 EU/mg and >99% purity.
Acknowledgment
Fig 4. 777 delivered 170 regular IgG with high titer, low endotoxin and high purity
We thank all scientists from WuXi Biologics’ Protein Sciences Department for their great support during the development of 777 platform. We also thank the legal department, public relations department and marketing department for their help in patent filing and poster preparation and revision. 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
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For more information see us at wuxibiologics.com
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To discuss this poster: PS_BD@wuxibiologics.com
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