We found a major flaw in a scientific reagent used in thousands of neuroscience experiments — and we’re trying to fix it.

[Jaybee00]

https://www.thetransmitter.org/open...cience-experiments-and-were-trying-to-fix-it/

 

[Jonathan Edwards]

Nothing new about the problem but it is good that someone has tried to address it systematically.

 

[Hutan]

The problem seems to be that antibodies used to identify proteins aren't as specific as people often like to imagine.

The C9ORF72 example illustrates a widespread problem. Antibodies are one of the most commonly employed reagents in molecular research, used to identify single proteins in a cell’s complex mixture. But scientists have known for decades that they can be inaccurate, binding to more than just the protein of interest. Publications that unknowingly use inaccurate antibodies can compound the issue, making it difficult to reproduce scientific results and raising questions about the validity of some preclinical drug studies.

Despite the seriousness of the problem, the field lacks a systematic way to characterize antibody accuracy. Quantifying how precisely an antibody highlights its target — its selectivity and specificity — is expensive and time consuming. The gold-standard approach is to compare cells expressing a target protein and those genetically modified to lack the target protein. Though many manufacturers do some knockout testing, the process is too expensive to apply to all antibody products. Most labs lack the requisite technologies, time or funding to rigorously characterize antibodies. As a result, most homemade or commercial antibodies are not subject to strict testing, a serious structural failure in the antibody ecosystem.

 

[Jonathan Edwards]

The problem seems to be that antibodies used to identify proteins aren't as specific as people often like to imagine.

And we have known that in spades since we started using monoclonals. I was the first person, as far as I know, in 1979, to use monoclonal antibodies to stain joint tissues in rheumatic disease. The problem was readily apparent from the start. And the reason was obvious. With traditional polyclonal antibodies the cross reactivities of all the component Ig species just gave a bit of background noise, which can be absorbed out with something like liver powder if needed. But for a monoclonal a single cross reactivity could easily be ten times stronger than the specificity you are wanting. You cannot absorb that out for a monoclonal because you just absorb out all your reagent.