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CRO Services

Drug Development Expertise Empowering Research Services for Biologics

Antibody Design & Engineering Services

Optimizing Antibody Developability Through Molecular Design and Lead Optimization

Our antibody design and engineering services integrate late-stage CMC insights into early drug discovery stage to enhance developability and manufacturability. By leveraging in silico developability assessments and advanced structural biology techniques, like X-ray crystallography, cryo-EM, protein modeling, and comprehensive analytical assays, we optimize critical attributes of lead molecules to achieve enhanced expression titer and minimized product-related variants (e.g., aggregates and fragments), off-target interactions, and immunogenicity risks by improving structure stability. Our advanced approach streamlines molecular design, lead optimization, and antibody engineering for monoclonal antibodies (mAbs), bispecific antibodies (bsAbs), antibody-drug conjugates (ADCs), and novel modalities.

 

Key Features of Antibody Design & Engineering Services:

  • In silico modeling for developability profiling, including solubility, pharmacokinetics, immunogenicity, and manufacturability
  • Comprehensive characterization through SPR binding assays, cell-based functional studies, and in vivo PK/PD studies
  • Tailored solutions to meet preclinical needs with expert data interpretation and troubleshooting

WuXi Biologics offers early-stage antibody design and engineering services, utilizing in silico assessments, structural characterization, and protein engineering to optimize developability and manufacturability profiles.

Antibody Design & Engineering Service Details

Service Item Turnaround Time Request A Quote 
Micro Developability assessment for antibody evaluation 1-2 weeks Request A Quote
In silico and structure-guided analysis for targeted antibody and protein engineering  1 week
Antibody & protein production for designed constructs 3-4 weeks
SPR/BLI binding characterization  1-2 weeks
Additional assays as needed  1-2 weeks

Case Study #1: Eliminating Bispecific Antibody Precipitation through Structural Analysis & Design

In this case study, we addressed agitation-induced precipitation of a bispecific antibody (bsAb) through in silico analysis and in vitro testing of charge distribution and hydrophobicity. The results identified conformational instability of antibody B as a key contributor to bsAb aggregation.

 

Structural mapping of Antibody B shows conformational instability contributing to bsAb aggregation.

Figure A: Agitation-induced precipitation and smart analysis revealed structural instability of antibody B.

Structure and sequence analyses identified two hydrophilic residues (VL_Ser7 and VH_Thr43) in the hydrophobic core of VL and the VH-VL interface, contributing to structural instability. Substituting these residues with hydrophobic counterparts improved the thermal stability of antibody B by 5.1 °C without affecting cell binding or function. The optimized antibody B, when combined with antibody A, yielded a bsAb without detectable precipitation at high concentrations.

Diagram showing Antibody B's enhanced thermal stability, FACS binding retention, reduced turbidity, and manufacturability improvements from optimization.

Figure 2: (A) Enhanced thermal stability of antibody B through sequence optimization. (B) In vitro FACS and reporter gene assays confirmed that the optimized antibody B retained its binding and functionality. (C) Turbidity analysis of the optimized bsAb revealed the elimination of precipitation, enabling further development through CMC activity testing. (D) This study has been published in Antibody Therapeutics.

Case Study #2: Resolving Bispecific Antibody Precipitation & Non-Specific Binding Issues by Structure Analysis and Rational Design

In this case study, a bsAb containing a VHH domain exhibited significant precipitation at low concentrations and high non-specific binding to Jurkat cells (Figure 1). High BVP ELISA scores (~30) and low homogeneity in DLS analysis indicated potential developability challenges, which were attributed to the presence of hydrophobic and positively charged patches, as confirmed by structural analysis (Figure 2).

ELISA and binding data showing bsAb precipitation and high non-specific binding to Jurkat cells.

Figure 1: Turbidity analysis of the bsAb at varying concentrations revealed precipitation. Additionally, FACS binding analysis demonstrated high non-specific binding to Jurkat cells.

DLS data, 3D model, and hydrophobic analysis of bsAb showing low homogeneity, positively charged patches, and aggregation-prone regions before optimization.

Figure 2: (A) DLS analysis highlighting low homogeneity. (B and C) Structural mapping identified positively charged patches and hydrophobic regions contributing to aggregation and non-specific binding.

Hotspot identification revealed critical regions contributing to developability issues. Through rational design, mutations were introduced to optimize surface charge and hydrophobicity. First, a single mutation at Site A significantly reduced the BVP score but did not improve homogeneity. Then, combining double or triple mutations targeting hydrophobic patches at Sites B, C, and D successfully eliminated aggregation and improved homogeneity (Figure 3).

Graph showing reduced BVP score and improved homogeneity, with reduced aggregation after surface charge and hydrophobicity optimization of bsAb.

Figure 3. (A) Charge optimization reduced BVP scores but did not improve homogeneity. (B) Optimization of hydrophobicity enhanced homogeneity and eliminated aggregation.

After optimization, the bsAb demonstrated significant improvements in solubility (increased from 11 mg/mL to 116 mg/mL) and elimination of non-specific binding while retaining functionality, as confirmed by in vitro assays (Figure 4).

Data showing increased solubility, reduced non-specific binding, and preserved functionality after bsAb optimization.

Figure 4. Optimization results demonstrated improved solubility, reduced non-specific binding to Jurkat cells, maintained binding affinity (Ka, Kd, KD), and preserved functionality, as evaluated by reporter gene assays.

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Frequently Asked Questions for Antibody Design & Engineering

Q: What is the optimal synapse distance for bispecific T cell engagers (TCEs)?

A: The optimal synapse distance is generally around 20 nm. However, this often requires extensive evaluation. That’s why our Quick ‘n’ Clean service is used for rapid production of different TCE formats for comparative analysis in T cell-mediated tumor killing and cytokine release assays, allowing us to identify optimal formats with maximal cytotoxicity and minimal cytokine release.

Q: How do you balance potency with manufacturability of bispecific antibodies to avoid redesign and timeline delays?

A: It is critical to incorporate developability assessments during early lead discovery. After small-scale expression, we recommend Micro Developability assessments, including BVP/DNA/Insulin ELISA, AC-SINS, DSF, and SEC. These help identify risks early. At the medium to large scale, our Premium BsAb production provides sufficient material for broader developability and in vivo studies. Based on our experience, it’s generally easier to improve potency through affinity engineering than to fix poor developability. While AI tools are emerging, early-stage experimental developability assessments remain critical.

Q: How do you guide bsAb design and format recommendation?

A: We apply rational design strategies. For example, scFvs are generally less stable than nanobodies or Fabs. To address light chain mispairing, we often use asymmetric designs where one arm is a nanobody or scFv and the other arm is Fab. Additionally, the WuXibody™ format replaces the CL/CH1 domains with TCR constant regions, which eliminates light chain mispairing. This format has shown strong performance and developability.

Q: Do you have experience designing bispecific antibodies that target two membrane proteins?

A: Yes, during antibody discovery, we can generate bispecific antibodies against any membrane-bound targets. For dual receptor targeting, the main challenge lies in the geometry of bispecific antibodies. Proper format design is crucial to enable simultaneous binding to both receptors on the same cell surface to effectively block signaling.

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