Bispecific Antibody Production Services | CRO Services

Integrated Discovery Service

Industry-leading expertise, state-of-the-art facilities, and multiple antibody generation technology platforms for the discovery of novel monoclonal, bispecific and multispecific antibodies, immunocytokines and other biologics.

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Protein Sciences

Specializing in monoclonal, bispecific, and multi-specific antibodies, fusion-proteins, cytokines, enzymes, and other biologics, we offer our customers high productivity and high quality antibody and protein production services for all of your research needs.

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Cell Line Engineering

Provided as either a standalone service offering or as part of our integrated CMC development platforms, WuXi Biologics provides extensive expertise and industry-leading timelines for cell line engineering and strain development across a wide range of biotherapeutics.

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Analytical Sciences

Our extensive analytical testing services offer top-tier expertise in method development for in-process, release, and stability assays, and support of cell line, process, and formulation development activities, product characterization, developability assessments and other key IND- and BLA-enabling studies.

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Process Development & Scale Up

Multiple upstream and downstream PD labs facilitate the establishment and scale-up of fed-batch, intensified fed-batch, and continuous manufacturing approaches for diverse biotherapeutic modalities in both early and late-stage development.

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Formulation & Drug Product Development

We provide advanced drug product development services spanning biologics, vaccines, and small molecules, with expertise in formulating clinical and commercial products in liquid, frozen, and lyophilized forms, compatible with various container closure systems such as vials, prefilled syringes, and other combination product types.

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Cell Banking & Characterization

Offering in-house, one-stop cell banking and cell line characterization services, compliant with global GMP regulations and ICH guidelines, we operate over 20 cGMP cell banking suites ensuring capacity availability and timely execution of this critical CMC development activity.

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Viral Clearance

Our extensive experience and expertise, along with EMA, ISO (CNAS), and CMA-certified laboratories, and exceptional track record for IND- and BLA-enabling viral clearance study acceptance by global regulatory agencies, form the cornerstones for this pivotal aspect of any CMC development program.

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Clinical DP GMP Manufacturing

Multiple, highly flexible clinical-scale drug product (DP) manufacturing facilities for the formulation, fill, labelling and packaging of biologics and parenterals under Current Good Manufacturing Practice (cGMP) conditions as defined by global regulatory agencies.

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Commercial DS GMP Manufacturing

We provide a global dual source manufacturing network that employs multiple large-scale drug substance (DS) GMP manufacturing facilities for the production of biologics utilizing our world-class operations and regulatory agency-approved quality systems.

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Commercial DP GMP Manufacturing

Multiple large-scale drug product (DP) manufacturing facilities located across the globe for the high-quality formulation, fill, labelling and packaging of biologics, parenterals and placebo into a wide array of container closure systems.

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Biosafety Testing

High quality, in-house and EMA, ISO (CNAS) and CMA certified biosafety testing facilities for adventitious agent screening of raw materials, cell lines and unprocessed bulk along with our extensive viral clearance capabilities is available to provide our clients with a single-source biosafety testing service offering.

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Global Quality Control (QC)

Regulatory compliant, in-house, QC labs support all clinical and commercial GMP sites pre- and post-production, ensuring top-quality testing for products and oversight of other critical functions (e.g., environmental monitoring, cleaning validations, instrument life-cycle and sample/retain management and audits).

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Global Compliance

Oversees global quality system and audit, IT quality, internal compliance, and risk analytics functions. This team embeds quality and compliance across the entire organization to ensure the products we manufacture are of the highest efficacy and safety standards and in line with your expectations.

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Quality Control

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Regulatory Affairs

Our experienced regulatory affairs team supports our client’s CMC dossier, drug filing and registration needs. Since 2015 this team has supported 550+ INDs, CTAs, BLAs, MAAs, NDAs and EUAs, and written 200+ Module 3 CMC packages on behalf of our global clients.

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Target to Lead

Providing expertise, highly efficient tailored solutions, state-of-the-art technologies and world-class facilities for the engineering, generation, and optimization of antibodies and other biologics from target identification to final lead selection.

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Bispecific & Multispecific Antibodies - WuXiBody™

The WuXiBody™ platform for the generation of bispecific (bsAb) and multispecific (msAb) antibodies speeds up drug development by 6-18 months, cuts production costs, and enables flexible assembly of mAb sequence pairs into various formats with different valency.

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Advanced Hybridoma Platform - WuXiHybrid™

Using advanced technologies to generate diverse monoclonal antibodies (mAb) for various targets. Our experienced scientists overcome technical challenges to deliver high-quality mAbs to our clients worldwide.

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Naive Phage Display Library - WuXiLiAb™

Offering high-quality phage display library construction and customized library selection and screening services via our WuXiLiAb® human naïve / synthetic phage display libraries, VHH naïve / humanized VHH synthetic libraries and antibody immune libraries.

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VHH-based Multispecific Antibody Platform - SDARBody™

Novel technology platform to enable the development of multispecific proteins. Highly flexible platform capable of generating multispecific antibody and protein therapeutics with high affinity, low immunogenicity risk and excellent developability characteristics.

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ScFv-based BsAb & MsAb Antibody Platform - SKYBody™

An innovative scFv (single-chain fragment variables)-based platform for the generation of bispecific or multispecific antibodies from nearly any sequence pair. The platform significantly improves scFv stability providing bsAbs and msAbs with strong developability characteristics.

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Ultra-Intensified Fed-batch platform - WuXiUI™

Ultra-intensified fed-batch bioprocessing platform utilizes an intermittent perfusion approach to achieve 3-6-fold higher productivity and 60 – 80% reduction in manufacturing cost of goods compared to traditional fed-batch processes.

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Rapid Research Material Generation - WuXian™

Custom protein generation services, producing high-quality, research-grade proteins/antibodies by using our industry-leading team of cell culture, purification and analytical experts and high-throughput, cost-effective production technologies.

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Continuous Bioprocessing - WuXiUP™

This ultra-high productivity continuous bioprocessing platform, achieves 5-15X greater productivity, delivering high-yield, quality drug products with flexibility and cost-effectiveness through intensified perfusion culture, continuous harvest and optional downstream continuous product capture step.

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High Concentration & HTP DP Platform - WuXiHigh™

Advanced high throughput formulation platform for development of high concentration biologics drug products using distinctive viscosity reducers and synergistic excipient combinations to enhance formulation stability and reduce viscosity.

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Scale-Out Biomanufacturing

Helping to pioneer this biomanufacturing strategy for higher volume production, we can help you reduce risk and increase manufacturing flexibility across the entire product lifecycle by using multiple, same-scale bioreactors to meet clinical and market demand.

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Robotic Aseptic Filling

Reducing manufacturing risk through the use of this fully programmable, robotic, gloveless, isolator-based technology for advanced aseptic drug product fill processing. These systems provide extensive flexibility to support complex fills across a wide range of biologics and container closure systems.

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Quick ‘n’ Clean: High-Throughput Bispecific Antibody Production

WuXi Biologics’ Quick ‘n’ Clean platform enables high-throughput bispecific antibody (bsAb) production for early-stage drug discovery. Powered by high-titer CHO expression system, it produces 1-7 mg of custom bispecific antibodies with 99% heterodimer purity from 20 mL CHO in just 3-4 weeks. Our fully automated workflow, integrated with high-throughput SPR binding assays, provides a rapid, cost-effective approach for optimal pairing identification. The bsAbs produced are not only suitable for in vitro screening assays, but also applicable for some in vivo studies.

Case Study #1: High-Purity Bispecific Antibody Production Across Various Formats

The production of bispecific antibodies typically involves a complex and time-consuming purification process to remove impurities caused by HC mispairing, LC mispairing, fragmentation, and aggregation. With our Quick ‘n’ Clean platform, the customized formats of bispecific antibodies, which comprise a scFv or VHH and a Fab on a heterodimeric Fc, can be purified in a high-throughput and automated manner, thereby eliminating the possibility of light-chain mispairing.

Case Study #2: High-Throughput Four-Chain Bispecific Antibody Production with the TFC Quick ‘n’ Clean Platform

Our True Four Chain Quick ‘n’ Clean platform offers high-throughput production of four-chain bispecific antibodies with excellent yield and purity. Leveraging the WuXiBody™ backbone, this advanced platform eliminates LC mispairing, enabling the production of bispecific antibodies with two LCs. This process ensures >99% heterodimer purity, as verified by Intact Mass analysis.

Premium Bispecific Antibody Production Services: LC-MS-Assisted Purification for High Purity

WuXi Biologics Premium Bispecific Antibody production services overcome key challenges in bispecific antibody (bsAb) production including chain mispairing, homodimer formation, and incomplete assembly. Leveraging ultra-high-titer CHO expression system, advanced LC-MS-assisted purification, and chain ratio optimization, this platform delivers high-yield, high-purity bispecific antibodies at milligram-to-gram scale within 4-6 weeks. With expertise in diverse bsAb formats, we optimize each stage of production to enhance efficiency, quality and efficacy.

Case Study #2: Removing Homodimers with Mixed Mode & CEX Purification for High-Purity Bispecific Antibodies

This case study presents a common LC bispecific antibody construct. After affinity purification, the HMW, half antibody and homodimer were observed based on SEC-HPLC, SDS-PAGE gel and Intact MS. Mix mode was used as the first polishing step. After that, most of the whole half antibody was removed. However, the homodimer still existed, indicated by LC-MS. CEX was applied as the second polishing step to eliminate the homodimer and trace amount of half antibody.

Frequently Asked Questions for Bispecific Antibody Production

Q: Do you use SEC for large-scale bispecific antibody production?

A: No, SEC is not ideal for large-scale bispecific antibody production due to scalability limitations. Instead, we utilize CEX chromatography or mixed-mode purification to efficiently remove aggregates in large-scale production. However, during early-stage antibody discovery and high-throughput antibody production, SEC remains a valuable tool for achieving high-purity bispecific antibodies for research purposes.

Q: Do you use LabChip instead of SDS-PAGE for bispecific antibody analysis?

A: The choice between LabChip and SDS-PAGE depends on throughput requirements. For high-throughput analysis, such as for 200 or more molecules, we use LabChip. For lower-throughput applications, SDS-PAGE is preferred due to its simplicity and ease of handling.

Q: Do you have experience with light chain mispairing? How do you address this issue (e.g., CrossMab)?

A: Yes, we have extensive experience in mitigating light chain mispairing in bispecific antibody engineering. To address this issue, we first optimize the light chain-to-heavy chain ratio to enhance proper pairing. If mispairing persists, it may indicate poor light chain expression. In such cases, we recommend swapping the VH-VL pair in the Fab arm with the VH-VL pair in the cross arm. Alternatively, switching the VH and VL domains within the same cross arm can further reduce mispairing.

Q: What recommendations do you have to generate heterodimers of four-chain bispecific antibodies?

A: We optimize chain ratios upon request to enhance the purity and titer of four-chain bispecific antibodies. While we have default expression conditions and extensive experience across various bispecific antibody formats, we strongly recommend conducting a chain ratio study, especially for large-scale bispecific antibody production, to achieve optimal yields. However, chain ratio optimization has its limits if one chain is poorly expressed. We suggest early-stage antibody engineering, sequence design, and micro developability testing to improve overall expression and stability.

Q: How do you decide between CEX, HIC, and mixed-mode chromatography for bispecific antibody polishing?

A: The selection of CEX, HIC, or mixed-mode chromatography depends on the pI differences and hydrophobicity of the molecule. CEX chromatography is preferred when there is a sufficient pI difference between product-related variants. If pI differences are minimal, HIC or mixed-mode chromatography is employed to enhance bispecific antibody purification efficiency. When light chain mispairing is a concern, HIC may offer better performance than mixed-mode chromatography.

Q: What purification steps do you use to ensure good viral clearance for WuXiBody™ given its low pI?

A: At the research stage, viral clearance is not a primary concern. However, during the CMC stage, viral clearance is addressed through acid treatment, exploiting WuXiBody’s tolerance to low pH, or by using detergents for molecules sensitive to acidic conditions.

Q: Do you use KappaSelect or LambdaFabSelect for Fab-scFv purification?

A: We generally avoid KappaSelect for Fab-scFv purification, as it can lead to aggregation issues, especially when handling large numbers of molecules in high-throughput antibody production. While LambdaFabSelect offers an alternative, it often exhibits low elution efficiency, making it less suitable for routine use due to potential bispecific antibody purification challenges. Instead, we employ optimized purification strategies tailored to the specific formats to achieve high purity and yield.

Q: Are Protein A Knockout Mutations Beneficial for Bispecific Antibody Purification?

A: In bispecific antibody (bsAb) production, Protein A knockout (KO) mutations can simplify purification by reducing byproduct binding to Protein A columns, which can be useful for early-stage in vitro studies. However, this approach is not recommended for in vivo studies, as the half-life of bsAbs with one Fc bearing the FcRn-null mutation has not been well understood.

Q: Why is LC-MS analysis so important during the purification of bispecific antibodies?

A: LC-MS analysis is essential for guiding bispecific antibody purification, as it accurately distinguishes heterodimers from homodimers, which often have similar molecular weights. Traditional methods like SDS-PAGE or size-exclusion chromatography (SEC) lack the resolution needed to differentiate these variants effectively. Alternatively, LC-MS analysis ensures high-purity bispecific antibody production, improving both yield and product quality.

Q: To produce bispecific antibodies, can I use my own tags, or do we have to use WuXi Biologics’ tags?

A: We typically recommend using our optimized tag system for bispecific antibody production, as it is designed to enable high-throughput purification, achieving high purity, and in a cost-effective manner. However, if you have preferred custom tags for your bispecific antibodies, we can evaluate their compatibility with our high-throughput antibody production platform and discuss potential integration to meet your specific needs.

Q: For small-scale bispecific antibody production, can the tags be removed afterwards?

A: For our Quick ‘n’ Clean platform, our primary aim is to generate ~1 mg of highly purified bispecific antibody in a fast and cost-effective manner to help unlock the optimal pairings of bispecific antibodies. The molecules don’t need to be in their final formats, so there’s no need to remove the tag. After the optimal pairings are identified, we can re-clone the chosen molecules into the final CMC-ready format and use our Premium Bispecific Antibody Production Service to generate the bispecific antibodies without a tag.

Q: Do you use antibodies to isolate tag antibodies? If so, are there concerns about anti-tag antibody leakage during purification?

A: Resin usage always poses potential leakage concerns. The key is to understand your molecule’s key quality requirement. For early-stage molecules, leakage isn’t often a significant concern. To our knowledge, we don’t see significant leakage of resin, even with our Quick ‘n’ Clean service. Those seeking extra assurance can opt for our Premium Bispecific Antibody Production Service in which we don’t rely on any tags and use Process Development (PD)-like resin to handle all steps of your purification to achieve very high quality.

Q: Do you use transient or stable transfection for the chain ratio study, and have you observed any discrepancies between the two transfection approaches?

A: Chain ratio studies inherently differ between transient and stable transfection approaches. Transient transfection allows for more extreme chain ratios, such as pairing Fc fusion with Fab at a 6:1 ratio, which can be challenging to achieve in a stable transfection system. However, since chain ratio reflects the molecular nature of the bispecific antibody, transient transfection chain ratio studies generally correlate well with stable pool chain ratio studies. Before transitioning to CMC development, we offer customized options based on project needs. Many clients prefer stable pools for chain ratio studies due to their scalability and consistency. Our bispecific antibody production services are flexible to accommodate specific preferences for your antibody discovery and development projects.

Q: Is the chain ratio optimization achieved by changing the ratio of input DNA constructs?

A: Yes, in our transient transfection system, we clone the heavy chain (HC) and light chain (LC) into separate vectors, allowing flexible adjustment of the input DNA construct ratio to achieve the desired chain ratio efficiently. For stable cell lines, we use molecular biology techniques to integrate multiple copies of the target molecules into one, two, or even three vectors. While this approach increases gene copy number, it offers less flexibility compared to transient transfection and involves a longer development timeline due to the complexities of molecular biology.

Q: Could the early-stage purification process be transferred to the CMC process?

A: Yes, absolutely. We work very closely with our Process Development (PD) teams to transfer our protocols, knowledge, and experience. For the developability assessment of bispecific antibodies in our PS Department, we use PD-ready resin to screen for different methods. These methods are amenable for scale-up and can be transferred to our PD department for further optimization. However, if your focus is solely on getting a large amount of high-quality bispecific antibodies without a developability study, some methods from our Premium Bispecific Antibody Production Service may not be readily transferred to the PD department.

Q: Do you offer a service to design bispecific antibodies?

A: We provide custom bispecific antibody design and engineering services, leveraging our extensive expertise in antibody engineering and drug developability assessment. Beyond simply designing bispecific antibody molecules, we evaluate various bispecific antibody formats to recommend the most suitable candidates for late-stage drug development and manufacturing. Our goal is to help clients identify molecules with the highest potential for therapeutic success, ensuring optimized bispecific antibody production with desired developability attributes.

More FAQs

Service Item	Deliverables	Turnaround Time	Quality Control	Request A Quote
Gene synthesis, cloning and plasmid prep all included.
20 mL	• 1-7 mg bispecific antibody • 3-step automatic purification • 99% heterodimer	3-4 weeks	<0.1 EU/mg endotoxin control, QC incl. Titer, A280, Caliper (R/NR) or SDS-PAGE, SEC-HPLC	Request A Quote
Additional QC	RP-HPLC, bioburden, DSF, DSC, SPR, LC-MS, SEC-MALS, peptide mapping, glycan profiling, cIEF, CHO HCP, residual DNA, ELISA, Western blot, freeze thaw study, micro developability testing

Premium Bispecific Antibody Production (Week)				Request A Quote
Week 1	Week 2	Week 3	Week 4	Request A Quote
DNA Synthesis and Prep	WuXian Transient™ Expression	Purification and Analysis
	Additional 2 weeks for ratio study	Analysis includes: LC-MS, SDS-PAGE, SEC-HPLC Purification scouting Purification scale up

High-Titer, High-Purity Bispecific Antibodies Across Diverse Formats

Quick ‘n’ Clean: High-Throughput Bispecific Antibody Production

Key Features of Quick ‘n’ Clean Service

Case Study #1: High-Purity Bispecific Antibody Production Across Various Formats

Case Study #2: High-Throughput Four-Chain Bispecific Antibody Production with the TFC Quick ‘n’ Clean Platform

Premium Bispecific Antibody Production Services: LC-MS-Assisted Purification for High Purity

Key Features of Premium Bispecific Antibody Service

Case Study #1: Optimizing Chain Ratio for Bispecific Antibodies for Higher Purity & Yield

Case Study #2: Removing Homodimers with Mixed Mode & CEX Purification for High-Purity Bispecific Antibodies

Frequently Asked Questions for Bispecific Antibody Production

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