The Case for Early Developability Assessment in Antibody Discovery
Scientist
on
December 9, 2025
This blog post was written by Biointron, a leading high-throughput recombinant antibody/protein expression and discovery service provider. Their services are available on Scientist.com.
… about 90% of drug candidates fail during clinical development, and this figure may exceed 99% when preclinical stages are included.
Therapeutic antibodies are driving innovation across a wide range of disease areas, such as oncology, autoimmune diseases, infectious diseases and neurological disorders. The pace of clinical development is accelerating, with over 570 antibody therapeutics in the clinical pipeline and dozens reaching late-stage development or approval each year. Despite this growth, the overall failure rate in drug development remains a significant challenge. According to published analyses, about 90% of drug candidates fail during clinical development, and this figure may exceed 99% when preclinical stages are included.
Antibody drug discovery revolves around functional performance: affinity, specificity and biological activity, as the principal criteria for lead selection. However, over the past decade, a growing body of evidence has underscored the importance of evaluating developability characteristics. Antibody developability refers to the physicochemical, biochemical and structural features that influence the manufacturability, stability and clinical suitability of a molecule. These properties can be the limiting factors that determine whether an otherwise promising antibody can advance through development.
In discovery-stage pipelines, delaying developability assessment until later stages can introduce significant risk. Candidates that perform well in binding assays may fail in formulation due to aggregation, instability or expression limitations. Early developability screening mitigates this risk by identifying such liabilities before costly downstream investments are made.
Early-Stage Developability Assessment: Scope and Purpose
Early-stage developability screening focuses on identifying and triaging candidates with unfavorable physicochemical properties, typically following hit identification or during lead optimization. Unlike chemistry, manufacturing and controls (CMC)-stage assessments, which require hundreds of milligrams of protein and weeks of testing, discovery-stage evaluations are designed to be rapid and efficient, using low material input and high-throughput formats.
Early-stage developability screening includes a set of widely accepted assays designed to identify major developability risks:
- Thermal Stability by DSF, nanoDSF: Measures Tm and Tonset to predict conformational robustness.
- Aggregation by SEC-HPLC, DLS: Detects soluble aggregates and self-association tendencies.
- Hydrophobicity by HIC-HPLC: Identifies surface-exposed hydrophobic regions linked to viscosity and instability.
- Charge Heterogeneity by iCIEF, IEX, Heparin HPLC: Determines pI and charge variants that affect purification and viral clearance.
- Non-Specific Binding by PSR ELISA: Measures polyreactivity to unrelated proteins such as insulin, BVP and/or DNA.
Charge-based assays such as imaged capillary isoelectric focusing (iCIEF), ion exchange chromatography (IEX) and heparin HPLC identify heterogeneity that may complicate purification, viral clearance or formulation stability. Additional tests assess non-specific interactions through poly-specificity reagent (PSR) ELISAs using DNA, insulin and other unrelated targets. These screens, while fast and relatively low-cost, are highly informative and predictive of downstream risks.
Differentiating Discovery-Stage and CMC-Stage Developability
Zhang et al. (2023) emphasize the critical distinction between early and late developability assessments. Discovery-stage evaluations aim to flag risk factors across a panel of candidates using minimal material, whereas CMC-stage assessments validate the manufacturability and long-term stability of a selected lead. At the discovery stage, even modest differences in Tm, aggregation propensity or charge profile can inform decisions on which candidates to prioritize, re-engineer or eliminate.
Meanwhile, CMC-stage assessments include extensive stress testing (thermal, oxidative, photolytic) and stability studies under multiple storage conditions. They also involve formulation screens with different buffers, excipients and concentrations, requiring several months. Thus, integrating developability assessments into discovery ensures that only structurally viable candidates advance to intensive downstream development.
Sequence-Encoded Liabilities Are Predictable and Actionable
Many developability risks are directly encoded in the amino acid sequence and are predictable using computational tools:
- Deamidation-prone motifs (NG, NS, NT): Can cause loss of binding or altered pharmacokinetics under stress.
- Oxidation sites (Met, Trp): Susceptible to degradation, especially when surface-exposed.
- Unpaired cysteines: Promote incorrect disulfide bonding and aggregation.
- Hydrophobic surface patches: Drive self-association and high viscosity.
- N-glycosylation sites in variable regions: Result in heterogeneity and potential immunogenicity.
In most cases, these liabilities can be mitigated early through conservative engineering. For example, replacing a deamidation-prone Asn with Gln or Ser can eliminate a degradation pathway. Removing an N-glycosylation site in the variable domain can improve product homogeneity. These modifications are far more efficient when performed prior to preclinical validation or lead selection.
Early developability profiling provides the necessary data to make such decisions confidently. By screening for risk motifs and testing for thermal and aggregation behavior empirically, discovery teams can implement a Quality by Design (QbD) approach from the outset.
Customizing Developability Criteria to the Intended Use
Developability is context dependent. The required solubility, viscosity and stability thresholds vary by indication, delivery route and formulation strategy. Subcutaneous (SC) delivery, for example, typically requires antibody concentrations above 100 mg/mL. At these concentrations, even modest self-association or charge imbalance can result in viscosity levels that exceed delivery device tolerances or patient comfort thresholds.
Intravitreal administration, used for ophthalmic indications, requires antibodies to remain stable at very high concentrations in extremely small volumes (e.g., 0.05 mL). Aggregation or charge variants in this setting can cause ocular inflammation or impaired bioavailability. For intravenous (IV) delivery, solubility constraints may be less stringent, but long-term stability and resistance to freeze-thaw degradation remain essential.
Antibody-drug conjugates (ADCs) and bispecific T-cell engagers also impose unique developability requirements, including high purity, minimal aggregation and low polyreactivity due to the risk of off-target toxicity or immune activation.
Defining the therapeutic context early allows for informed threshold setting during screening. By aligning developability assessment with intended use, teams can eliminate candidates that may be otherwise viable for a different modality, reducing late-stage reformulation or re-engineering.
Early Profiling Improves Development Efficiency
Developability assessment is most effective when applied immediately after hit generation or in parallel with functional screening. Candidates with acceptable binding and specificity profiles can be triaged based on developability performance, using quantitative metrics for direct comparison. For example, among two leads with similar antigen-binding kinetics, the molecule with a higher Tm, lower HIC retention and fewer charge variants is more likely to succeed in manufacturing and formulation.
Rather than acting as a binary pass/fail filter, developability data should be used to rank candidates and guide downstream efforts. Molecules with borderline characteristics may be reformatted, engineered or moved into alternative delivery strategies, while top-performing antibodies can be prioritized for scale-up.
High-throughput platforms make this approach feasible even for large antibody panels. Assays can be conducted with less than 1 mg of purified material, and results can be obtained within a few days. This enables efficient decision-making without adding significant time or cost to discovery timelines.
Biointron’s Developability Platform: Supporting Early-Stage Decision Making
Biointron offers a comprehensive developability assessment service specifically designed for early-stage antibody discovery. With the capacity to express over 3,000 antibodies per batch, we integrate expression, functional characterization (affinity, epitope binning, cell binding) and developability profiling into a streamlined workflow. This allows rapid assessment of both binding performance and drug-like properties in parallel.
| Analysis | Developability Factors | Timeline |
|---|---|---|
| SEC-HPLC | Protein quality | 3-5 days |
| CE-SDS | Protein quality | 3-5 days |
| DSF Tm & Tonset | Thermal stability | 3-5 days |
| AC-SINS | Self-interaction | 3-5 days |
| HIC-HPLC | Hydrophobicity | 3-5 days |
| DLS | Tagg/Radium analysis | 3-5 days |
| Heparin HPLC | Positive charge tendency | 3-5 days |
| IEX-HPLC | Charge heterogeneity | 3-5 days |
| iCIEF | Charge heterogeneity | 3-5 days |
| PSR ELISA | Non-specific Binding | 3-5 days |
Learn more about Biointron’s Antibody Developability Assessment → https://www.biointron.com/antibody-developability/antibody-developability-assessment.html
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