YCharOS: A Partner in the Fight for Better Antibody Validation and Characterization

As part of our commitment to supporting scientists at every level, Aviva Systems Biology has joined industry-wide efforts to overcome the reproducibility crisis observed in biomedical research plaguing researchers today. Importantly, our Enhanced Validation framework extends beyond internal efforts to improve reproducibility—a partnership with YCharOS, an initiative dedicated to antibody characterization and validation, is a vital aspect in our efforts.

YCharOS (Antibody Characterization through Open Science) is a non-profit organization aiming to improve antibody reliability and reproducibility in biomedical research by working directly with industry partners. The initiative is operated through the Agora Open Science Trust, a Canadian charity committed to advancing biomedical innovation through open science. YCharOS provides extensive, openly shared characterization data of antibody reagents available from many leading vendors, including Aviva. Its neutral, vendor-agnostic approach positions YCharOS as a trusted source within the research community.

Comprehensive Antibody Validation Standards

While antibodies are essential to many research techniques, inadequate validation of antibody reagents is a major contributor to persistent issues with the lack of reproducibility . Case in point, a recent evaluation of > 600 antibody products found that more than 50% failed in one or more experimental applications.¹

Validation of antibodies for their intended target and application is necessary but labor-intensive, leading many researchers’ validation efforts to fall short. Additionally, the lack of consensus over how to validate an antibody further complicates the path to adequate, standardized validation practices.² Testing antibodies against purified or overexpressed proteins is one common approach, but because endogenous levels are typically lower, this does not necessarily provide relevant data on an antibody’s selectivity or sensitivity. Conversely, genetic knockout (KO) cell lines or animals are ideal controls for antibody target validation but are not readily available for most proteins.  Generating these tools is expensive and often technically challenging, limiting accessibility for most researchers.

Recognizing this challenge, YCharOS was created to systematically characterize commercial antibodies against human proteins in common applications using rigorous, consistent, and transparent methods. The consensus protocol, developed through collaboration between industry leaders and researchers at the Montreal Neurological Institute (McGill University), was published last year in Nature Protocols.³ The process begins by identifying a cell line that can maintain relevant expression levels of the endogenous RNA/protein of interest, then generating a KO cell line using CRISPR-Cas9. YCharOS researchers then characterize the performance of multiple commercial antibodies from partner vendors side-by-side across western blot, immunoprecipitation and immunofluorescence.

Making an Impact Across the Industry with Open Data

All characterization data are made openly available on Zenodo and F1000, enabling researchers across every level of academia and industry to access these valuable insights into antibody reagent performance. YCharOS has published reports characterizing roughly 1,500 antibodies against over 150 protein targets. Their validation efforts primarily focus on high-value targets with relevance to critical areas of disease research, including neurodegeneration, cancer, and inflammation, to help researchers generate reliable and trustworthy experimental results.

In addition to its depth of analysis, the breadth of industry collaboration behind YCharOS is a defining factor in its success. This broad reach enables researchers to directly compare antibodies between different vendors. Through its 11 industry partners, including Aviva Systems Biology, YCharOS has access to approximately 80% of all commercially available recombinant and monoclonal antibodies, paving the way for its ever-expanding characterization efforts.

This collaboration does not just benefit researchers—it is a positive step forward for the entire industry, fostering enthusiastic investment in open science. Industry partners unite with YCharOS researchers for a monthly call to review recent data, discuss points of interest, and plan for the future. Vendor partnerships with YCharOS demonstrate an ongoing commitment to providing high-quality, reliable research tools and driving transparency and accountability in the antibody marketplace.

Our Shared Commitment to Scientific Transparency and Reproducibility

As an organization dedicated to transparency and quality, Aviva Systems Biology is proud to be among YCharOS’s industry partners. Aviva has independently adopted Enhanced Validation methods that provide customers with validation data for antibody products, including providing any genetic KO validation done through YCharOS on our product pages. Because polyclonal antibodies (pAbs) are often known to underperform, we proactively submitted multiple pAb products for YCharOS assessment. This validation process revealed strong performance for many of these pAbs, providing customers with clear and trustworthy validation data. More importantly, the data has helped identify poor or ambiguous performers that need to be reevaluated and potentially removed from the market.

Above all, Aviva Systems Biology works to provide researchers with well-characterized tools that help drive progress across the biomedical sciences. In addition to conventional validation methods and our continued partnership with YCharOS, we leverage the Carterra® LSA™ surface plasmon resonance (SPR) platform for powerful, high-throughput characterization of our growing recombinant antibody pipeline. Aviva also offers SPR antibody characterization as a service, providing other researchers and antibody developers access to the valuable kinetic data generated using this method.

Through our growing catalog of recombinant antibodies, support for open-access validation data, and high standards for antibody validation, Aviva Systems Biology aims to operate at the forefront of scientific rigor and reproducibility to empower every customer in their research.  

References

1. Ayoubi, R., Ryan, J., Biddle, M. S. Scaling of an antibody validation procedure enables quantification of antibody performance in major research applications. eLife 12, RP91645 (2023).
2. Baker, M. When antibodies mislead: the quest for validation. Nature 585, 313–314 (2020).
3. Ayoubi, R., Ryan, J., Gonzalez Bolivar, S. et al. A consensus platform for antibody characterization. Nat Protoc 20, 1509–1545 (2025). https://doi.org/10.1038/s41596-024-01095-8.