Q&A Part 6: Who Is Responsible for Antibody Reproducibility?

Rockland believes that both antibody manufacturers and researchers are responsible for validating antibodies. All antibody manufacturers should validate every lot of antibody produced by defined minimum acceptable criteria. The validation results, the details pertaining to the nature of the immunogen, and the screening strategies should be communicated to the researcher for transparency purposes. However, it is recommended that all researchers should qualify antibodies used in their laboratories before collecting experimental data to ensure expected performance using defined protocols, buffers, personnel, and equipment.

BD believes antibody validation is critical to ensure accurate scientific results. Both the consumer and the reagent provider share the responsibility for reproducible science.

It is our responsibility to validate the antibodies we commercialize. We must ensure that they recognize the correct target, do not recognize undesired targets, and we must provide as much context as we can regarding their optimal use. When we provide an antibody conjugate (e.g. fluor, enzyme, oligo) we must have proven that it will work readily in the application(s) we suggest. This is true for both the initial validation of a new antibody, and for each subsequent lot produced.

Ensuring scientific results are reproducible is the responsibility of both suppliers and end-users. Suppliers must ensure they are providing a dependable product. End-users must ensure they obtain their experimental materials from reliable suppliers, and then use such products in well-designed experiments that make appropriate use of the materials.

When scientists are validating an antibody for use in their experiments, they should:

• Always follow manufacturer’s recommendations
• Employ a relevant model system to test the antibody
• Use ample controls, both for the target (e.g. negative cell line) and for the reagent (e.g. isotype controls that are modified in the same fashion as your antibody)
• Compare to other similar antibodies if available
• If something looks amiss, contact the supplier’s Technical Service and share your data with their scientists.

At CST, no one group is solely responsible for ensuring the scientific rigor as there are many stakeholders involved. Suppliers should be held accountable for the products they provide and should be transparent in the source and validation of the products they sell. Vendors should also be held accountable for supporting and continually validating their reagents, as well as working with customers to improve, test, and support their products. Researchers need to remember that antibodies, unlike chemicals used in the lab, are dynamic, meaning sensitive to the context and method in which they are used. Researchers also need to leverage the expertise of the scientists who have developed and tested the antibody and carefully follow established protocols. A manuscript lacking sufficient identification of reagents and materials is, by definition, irreproducible. Therefore, journals need to actively enforce existing policies regarding materials and methods or develop definitive means to identify and describe the use of biological reagents in published research. While tools are being developed to facilitate this process (www.protocols.io, Research Resource Identifiers, etc.), the researchers performing the experiments and writing the papers are ultimately accountable for ensuring their work can be reproduced by others.

• At Miltenyi Biotec we think that all stakeholders are ultimately responsible for reproducibility. As an antibody manufacturer we see our primary responsibility in the development and implementation of consistent manufacturing processes and rigorous validation of the antibodies for their intended applications. This also means providing rigorously validated application protocols.
• By using recombinant antibody technology instead of hybridoma technology, we have eliminated many causes for lack of reproducibility in antibody production.
• We have recently launched campaigns on knockout validations (specificity), clone comparisons (sensitivity), and lot-to-lot comparisons (reproducibility) to provide our customers with even more antibody validation data.
• Apart from that we also offer hands-on trainings for our customers in our MACS^® Academy in order to make sure antibodies are used correctly in their target applications.
• Furthermore, we are a proud sponsor of the GBSI Antibody Validation Workshop as well as the the 2nd and 3rd International Antibody Validation Meetings.

Although there are no established and uniformly accepted requirements for antibody validation, it is of utmost importance that manufacturers provide details of their validation process and their data supporting the selectivity, sensitivity, and application specificity of their antibodies. Principal investigators should make sure that researchers working in their laboratory are sufficiently trained to handle these antibodies and conduct needed experiments following established protocols and obtain reproducible results. Attention should also be given to sample preparation to avoid loss of or degradation of proteins or their dephosphorylation that may be critical in some applications. In addition, experiments must include controls.