High-throughput, high-titer CHO production of miniproteins with high yields and low endotoxin levels, surpassing traditional E. coli production methods.
Bispecific Antibody Discovery: From Strategic Design to Fit-for-Purpose Screening & Characterization Macy Jin, Huihui He, Xiwen Ji, Jiameng Zheng, Donghui Wu, Gaozhan Zhang, Guoyun Zhu, Zhili Yao, Jiansheng Wu
WuXi Biologics, No.240 Hedan Road, Pudong New District, Shanghai, China (Contact: PS_Marketing@wuxibiologics.com)
BsAbs represent a versatile therapeutic class with diverse applications in T-cell engagers, bispecific ADCs, immune checkpoint inhibitors, signaling modulation in inflammatory pathways, blood brain barrier delivery, half-life extension, etc. They present significant opportunities to enhance therapeutic e ff icacy and specificity, overcome drug resistance, incorporate multiple mechanisms of action, and reduce treatment costs. However, bsAb discovery also encounter challenges in each stage. At WuXi Biologics, our practical discovery strategies encompass rational molecular design, antibody discovery platform selection, and fit-for-purpose screening and pharmacology characterization. We also emphasize critical aspects of TCE optimization, including epitope selection, a ff inity tuning, and PK assessment, to advance candidates with the optimal balance of e ff icacy, safety, and developability. Introduction Abstract Bispecific antibodies (BsAbs) enable diverse therapeutic MOAs such as retargeting cytotoxic e ff ector cells, modulating receptor activity, and selectively recognizing pathogenic cells. However, advancing bsAbs from concept to preclinical candidates remains challenging, requiring comprehensive insights and technical expertise at every stage. At WuXi Biologics, we have established comprehensive bsAb discovery strategies that streamlines rational molecular design, biology-driven antibody discovery platform selection, and fit-for-purpose screening and characterization. We further highlight key considerations for bsAb optimization like format and modality selection, high-throughput (HTP) production for optimal pairing identification, and tailored pharmacology assessments. Together, these capabilities facilitate a seamless transition into CMC development.
Fig.4 HTP protein- and cell-based binding assays (ELISA, BLI, SPR, and flow cytom- etry) to evaluate binding activity
Various Methods • Protein-based: • ELISA • BLI • SPR • Cell-based: • Cell ELISA • Flow cytometry
Multiple Cell Lines in ONE Well
Cell line mixture
1000 2000 3000 4000
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0.0001 0.01 1 100 10000 0
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[Abs]nM
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200 400 600 800
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Binding on a tumor cell line by FACS Binding on a tumor cell line by FACS
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hIgG1 Isotype Control
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Ab1 Ab2
Comp-FarRed-A
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0.00010.001 0.01 0.1 1 10 100 1000 0
Abs (nM)
This figure highlights one-stop approach to bsAb discovery, combining rational design with high-throughput binding assays to rap- idly screen early hits, providing early insights into format and modality selection.
Fig.5 TCE discovery workflow containing antibody binning for pairing, HTP produc- tion with Quick ‘n’ Clean, and in vitro activity assays
Fig.1 Comprehensive antibody discovery platforms with proven track of successful projects
Quick ‘n’ Clean enables high- throughput produc Ɵ onof TCEs
Pair TAA an Ɵ bodies from di ff erent air TAA an bo bins with an Ɵ od - s fromdi eren dies -CD3an Ɵ body
Invitro ac Ɵ vity assay of TCEs
Titer
Bin12
0.0 0.5 1.0 1.5 2.0 2.5 3.0
SingleB Cell Cloning
VHH Libraries Immune, Naïve & Synthe Ɵ c
1.92g/L
Rodent Hybridoma Human Ab Libraries Naïve & Synthe Ɵ c
Bin3
TCE Killing assay
Bin11
100
Lead 1 Lead 2 Lead 3 Lead 4 Lead 5 Lead 6 Lead 7 Lead 8 Lead 9 Lead 10
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350+ Successful projects
100+ Successful projects
100+ Successful projects
100+ Rodent,camelid & rabbit an Ɵ bodies
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Anti-Flag
CONC(nM)
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This figure demonstrates a streamlined workflow from antibody binning to HTP production and functional assessment, en- abling rapid identification of optimal TCE molecules.
Fig.6 Selected in vitro and in vivo studies evaluating TCE candidate function and e ff icacy
Fig.2 Cutting-edge lead engineering platforms
T Cell Mediated Tumor Cell Killing
NHP PK/PD Studies
Humanization
A ff inity Maturation · High success rate ݠ in improving a ff inity by 10-1,000x · Fast: ~3-4 months
Fc Engineering · E ff ector function (AD- CC,CDC) enhanced or eliminated · Half-life extended or shortened
Developability · Remove PTM hotspots · Improve expression · Improve solubility · Improve stability
· Fast: ~4 weeks · Success rate >98% · Applicable to rodent, camelid, rabbit and chicken antibodies
P329
L234
L235
N297
H435
T256
Y407 L368
· Both antibodies demonstrated potent tumor cell cytotoxicity. · Ab1 induced lower cytokine release compared to Ab2, suggesting a potentially improved safety profile.
NHP studies revealed a unique PD profile where PD e ff ects were not directly proportional to PK responses.
S254
M252
T366
Y436
S354
Conclusions WuXi Biologics provides a one-stop, modular solution for bsAb discovery, backed by extensive drug development expertise. Through biology-driven antibody discovery, rational molecular design and engineering, HTP screening and production, and comprehensive in vitro and in vivo pharmacology studies, we deliver high-quality candidates optimized for e ff icacy, safety, and developability. We also demonstrate how this approach streamlines lead identification and optimization, supporting e ff icient progression into later development stages. Acknowledgment We extend our gratitude to all scientists from WuXi Biologics CRO Services for their invaluable support during the development of the antibody discovery platform. Additionally, we express our appreciation to the legal department, public relations department, and marketing department for their assistance with patent filing, poster preparation, and revision. About WuXi Biologics CRO Services 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 antibody discovery, protein production and engineering, and in vitro / in vivo testing, to seamlessly advance our clients’ molecules from concept to CMC development.
Results
Fig.3 Strategic design of TCE format and modality
BsAb formats are key to the potency and cytokine windows of TCEs ： · Valency · Geometry · Synapse distance · PK · Developability
Anti-CD3 · Epitope · Cyno x-reactivity · Kinetic a ff inity
Anti-TAA · TAA selection · Epitope · A ff inity and avidity
Drug Development Expertise Empowering Research Services for Biologics For more information see us at https://www.wuxibiologics.com/cro-services
Antibody Discovery_Poster_202510_V3
To discuss this poster: PS_Marketing@wuxibiologics.com
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