Antibodies are central to modern biological research, yet inconsistent performance continues to challenge reproducibility across techniques like Western blot, immunohistochemistry, and ELISA.
At Aviva, we’ve adopted an Enhanced Validation framework to help address this industry-wide challenge. These validation strategies are being retro-applied to our catalog products for further confidence in performance. But importantly, these strategies are the backbone of our ongoing recombinant antibody manufacturing pipeline, providing robust evidence of antibody performance before a product even reaches your bench, saving time, improving data quality, and enabling more confident decisions.
The Reproducibility Crisis: Why Enhanced Validation Matters
Reproducibility is a cornerstone of science, yet more than 50% of life science researchers report difficulty replicating published results. A major factor is poor reagent quality, particularly unvalidated or under-validated antibodies. Inconsistent antibody performance leads to wasted resources, misinterpreted data, and delays in discovery.
Publishers and funding agencies are increasingly calling for validated reagents. Researchers now face growing pressure to choose antibodies that come with comprehensive, transparent performance data. Aviva’s Enhanced Validation program is built to meet that need.
Enhanced Validation vs. Industry Standard: What’s the Difference?
Aviva’s Enhanced Validation goes beyond minimal testing to deliver multidimensional evidence of antibody performance.
Genetic Validation: Definitive Evidence of Specificity
Genetic models remain the most trusted method for determining antibody specificity. In knockout systems, the complete loss of signal in a sample lacking the target gene offers clear evidence that the antibody is binding to the intended protein. Knockdown models, where gene expression is reduced rather than eliminated, offer additional insights and analysis.
Aviva is a proud partner of YCharOS, an independent initiative using genetic tools to assess commercial antibodies. See this approach in action with our TIA1 antibody, validated in knockout samples.
YCharOS (Antibody Characterization through Open Science) is a nonprofit organization dedicated to improving antibody quality across the life sciences. Through a collaborative and transparent approach, YCharOS systematically characterizes commercial antibodies using CRISPR-generated knockout cell lines. Each antibody is tested in multiple applications, including Western blot, immunofluorescence, and immunoprecipitation, to determine specificity and reliability. Their open-access model allows researchers to evaluate antibody performance based on independent, standardized data. Aviva contributes antibody candidates to this effort and uses the resulting validation data to strengthen product confidence and guide continuous improvement. See Aviva’s Press Release about YCharOS’s Published Consensus Protocols.
SPR Affinity Characterization: Quantifying Performance
Surface plasmon resonance (SPR) is a label-free, real-time technology used to study molecular interactions. In antibody validation, it plays a critical role by providing direct measurements of how strongly and how specifically an antibody binds to its target. SPR determines kinetic parameters such as the association rate (kon), dissociation rate (koff), and the equilibrium dissociation constant (KD), which together describe the binding affinity and stability of the interaction.
Aviva uses the Carterra® LSA™ platform for high-throughput SPR, enabling simultaneous analysis of dozens to hundreds of antibodies in parallel. This advanced technology not only accelerates our validation timelines but also ensures consistency in how binding data is generated and interpreted. The Carterra system supports both epitope binning and kinetic screening, making it ideal for developing and qualifying antibodies intended for therapeutic, diagnostic, or high-performance research use.
We conduct SPR as part of our Enhanced Validation pipeline to ensure that our antibodies meet stringent performance criteria, especially when binding affinity and specificity are critical to experimental success.
To learn more about how Aviva applies SPR in antibody validation, explore our SPR characterization capabilities or watch our co-sponsored 2023 webinar with Carterra, which details our methodology and results in depth.
Biological Expression: Target Verification in Context
To confirm specificity in biological systems, we evaluate antibody performance in cell lines and tissues known to express—or not express—the target protein. Consistent signal in positive models and absence of signal in negative models helps ensure the antibody is detecting the intended target, not cross-reacting with unrelated proteins.
Great examples of differential expression in our validations include antibodies against:
Independent Recognition: Cross-verifying with Multiple Antibodies
Another layer of confidence comes from using two or more antibodies that target different epitopes of the same protein. When these antibodies yield consistent results—particularly in Western blotting—it strengthens the evidence for specificity and minimizes the risk of misleading data due to off-target binding.
When developing our new recombinant antibodies, we also leverage epitope binning using the Carterra® LSA™ platform to map epitope diversity among our antibody panels. Epitope binning allows us to classify antibodies based on their binding sites on the target protein, enabling the identification of non-overlapping, independently recognizing clones. These interactions are visualized using community plots, which group antibodies into bins based on competitive binding patterns. These plots provide a powerful visualization tool to assess the uniqueness and redundancy of binding across a large panel of antibodies.
In addition to supporting validation and characterization, this capability allows us to identify complementary antibody pairs for use in sandwich immunoassays. By selecting antibodies that bind distinct, non-competing epitopes, we can help ensure assay sensitivity, specificity, and performance across detection platforms.
Our BDNF antibodies, for example, target the N-terminal, middle, and C-terminal regions of the protein, allowing researchers to cross-validate results using multiple reagents or build robust paired-antibody assays.
Looking Ahead: The Future of Validation
As the life sciences community moves toward higher reproducibility standards, antibody validation will only grow in importance. The shift toward recombinant antibodies, support for open data initiatives like YCharOS, and growing expectations from journals all point toward a more transparent future.
At Aviva, we’re committed to being part of that progress, by raising the standard for antibody validation and delivering the high-quality, reliable tools researchers deserve.
Look for the Enhanced Validation badge across our catalog, and reach out if you'd like to discuss the validation data behind any specific antibody.
