Antibodies are among biologists’ most essential tools, but many just don’t work. Worse, many researchers who rely on these protein-binding reagents don’t know the pitfalls. According to YCharOS, a Canadian company dedicated to open science, most commercial antibodies fail to recognize their target proteins or bind off-target in at least some experimental applications — squandering time and money and muddling research.
The solution? There may not be one. But there may be many. If scientists continue to talk about the issues and share ideas, the community as a whole may find better answers.
Earlier this month, the Institute for Protein Innovation (IPI) was part of one such conversation at “Antibody Validation: A Call to Action,” a sponsored session hosted by YCharOS at Cell Bio 2023, the joint meeting of the American Society for Cell Biology (ASCB) and European Molecular Biology Organization (EMBO). Rob Meijers, senior director of the Antibody Platform at IPI, participated in a panel discussion along with other academics, antibody manufacturers and audience members who added relevant insights.
Cleaning up the antibody tool chest will require efforts on all sides, discussants agreed. Here’s a look at some of the initiatives and ideas put forth:
- YCharOS is characterizing commercially available reagent antibodies for human proteins. The company uses cell lines that endogenously express target proteins, comparing them with knockouts. The initiative recently reported a first dataset — and, as first author Riham Ayoubi shared in the session, a substantial fraction of antibodies performed poorly.
- The high failure rate means researchers must investigate. “I’m amazed at the number of scientists who simply buy an antibody and assume it works,” said panel moderator Rick Kahn, a professor at Emory University School of Medicine.
- Harvinder Virk, a clinical lecturer at the University of Leicester in the UK and co-founder of Only Good Antibodies (OGA), wants to empower antibody users to proceed with caution. His appeal: Before you use an antibody, seek data. Perform appropriate validation tests yourself. When you publish results, identify your antibodies with Research Resource Identifiers (RRIDs). And share what you know. If an antibody doesn’t work, tell the provider, and submit your data to Biomed Resource Watch. (Read Virk’s personal story of “bad antibodies” in eLife.)
- Antibody manufacturers say they’re ready to field inquiries. Panelists from Abcam, Cell Signaling Technology, GeneTex and Thermo Fisher said their companies perform validation tests, but many details on the antibodies aren’t provided. The representatives encouraged researchers, before ordering, to reach out with questions.
- IPI’s Rob Meijers championed the open sharing of antibody data. IPI is generating synthetic recombinant antibodies for distribution through Addgene. “For us it’s really important also to share the sequences of these antibodies,” he said.
- Alongside antibody makers and users, funders and journals can play key roles in promoting effective antibody use. From the audience, Andrea Marat of the Journal of Cell Biology said she’d like for journals to start asking for antibody validation data. She noted that editors screen figures for signs of image manipulation — “but we don’t know if the antibody is good to begin with.” So she encourages reviewers to scrutinize and ask for validation data. “If reviewers start routinely asking,” she said, “I think that will help put pressure on the journals.”
- Audience members also brought up valid concerns. Researchers are busy, and validating antibodies takes time and money, noted Pina Colarusso of the University of Calgary. Colarusso urged manufacturers to incentivize researchers to validate with rewards such as low-cost samples.
- Doug Houston, director of the Developmental Studies Hybridoma Bank (DSHB) at the University of Iowa, also invited researchers to think beyond the antibodies that now exist: “Don’t be afraid of making your own antibodies.” A great new tool, he said, can transform a field.
For more on improving antibody reliability and accessibility, watch or read about the panel discussion at our June 2023 IPI Surfacing symposium.
Writer:
Megan Talkington, megan.talkington@proteininnovation.org
Caitlin Faulds and Trisha Gura contributed to this story.
About IPI
The Institute for Protein Innovation is pioneering a new approach to scientific discovery and collaboration. As a nonprofit research institute, we provide the biomedical research community with synthetic antibodies and deep protein expertise, empowering scientists to explore fundamental biological processes and pinpoint new targets for therapeutic development. Our mission is to advance protein science to accelerate research and improve human health. For more information, visit proteininnovation.org or follow us on social media, @ipiproteins.