A reliable method for characterizing antibody quality is reported in a paper published online this week in Nature Methods.
The reproducibility of scientific results is a major concern in biomedical research. In particular, problems with the reliability of antibodies have spurred efforts to improve the quality of these widely-used protein binding reagents.
Aled Edwards and colleagues describe a mass spectrometry-based standard operating procedure to score the performance of antibodies in immunoprecipitation experiments, in which an antibody is used to specifically bind and isolate a target protein from a complex mixture. They applied the method to more than 1,000 antibodies targeting 152 human proteins, classifying the antibodies as ‘gold standard’ (meaning the intended target protein was the most abundant protein isolated), ‘inconclusive’ (meaning the target protein was isolated but was not the most abundant protein) or ‘failed’ (meaning it did not isolate the target protein at all). Researchers in five independent laboratories tested the performance of the standard operating procedure and showed that antibody classification by this method was reproducible.
The authors suggest that their method could be applied in a community-based effort to benchmark antibodies. They call for every antibody intended as a biomedical research reagent to be accompanied by a publicly available package of mass spectrometry-based characterization data.