Scientists expose gaps in protein detection tests and suggest solution:
A new study highlights the need to create a better method for validating antibodies, and describes a process that would allow laboratories to ensure the proper functioning of their own.
In the laboratory and in the clinic, antibodies are used to study proteins, biomolecules which are the result of the translation of the genetic information of an organism and which play a role in the structure, function and regulation of its tissues. and organs. Genetic mutations can lead to protein imbalance or dysfunction, which can lead to disease.
Using antibodies, scientists can determine where and how much of a given type of protein is in the cell. Because of their usefulness, many companies manufacture antibodies for sale to research laboratories and clinical laboratories.
However, studies have shown that many antibodies already on the market do not specifically detect the proteins they are supposed to target. This finding calls into question all the studies that have used it, and demonstrates why it is necessary to develop a precise and standardized method to assess the quality of antibodies.
Under the direction of Peter McPherson, a team from The Neuro (Montreal Neurological Institute and Hospital) decided to use a human protein as an example, both to confirm the problem of antibody validation and to demonstrate the effectiveness of its new validation process.
The team chose the protein product of the C9ORF72 gene , whose mutations are the main genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia.
After testing 16 antibodies supposed to specifically detect the C9ORF72 protein, she discovered that only one detected it precisely with immunofluorescence, a technique that stains proteins to make them visible under a microscope. Only two antibodies have shown convincing results using other techniques. Antibodies that met the validation criteria have not yet been used in scientific studies, while the others have been used many times.
These results call into question the studies which have used to detect C9ORF72 antibodies which do not recognize the protein or which wrongly detect other proteins. They also highlight the need to improve validation methods.
Science is currently facing a crisis: the results of numerous studies have proven to be impossible to reproduce, and the lack of effective antibodies contributes to the problem in cell biology. In an article published on October 15, 2019 in the open access journal eLife , the authors describe their validation method, which other laboratories can reproduce to verify the effectiveness of their antibodies.