In the late 1970s, I led a research group at the Balearic Islands University to investigate metabolic changes during development. Since the university was new, we had a limited budget and no controlled animal facilities for rat experiments. 

Chick embryos emerged as a viable alternative. They were also an interesting model system as questions remained about the biochemical adaptations during early chick development, including the rate and location of protein synthesis.           

We addressed these questions by injecting fertilized eggs with carbon-14 labeled amino acids. After 24 hours of incubation, we dissected different egg components and measured the radioactive signal using a scintillation counter. 

The marked amino acids seemed to incorporate into proteins, indicating a rapid metabolization, which contrasted with the fact that protein synthesis is relatively slow. We also consistently found the labeling in the yolk and albumen, two components presumably not subjected to active synthesis by the embryo. Although we could not explain the findings to our complete satisfaction, we submitted them for publication and kept looking for explanations. Then, we found a paper that described how amino acids that appeared to be incorporated into proteins were actually physically attached, or adsorbed, to them.¹ 

We tested the adsorption hypothesis and found that to be the case in our experiments as well. Although we were shocked to discover that our interpretation of the results was incorrect, we felt satisfied that we could fully explain them. This feeling, though, rapidly turned to concern as we had just received the journal’s acceptance letter. It was unacceptable for us to publish what we knew to be wrong, so we quickly wrote to the journal’s editor asking to withdraw the manuscript. 

This experience showed us the value of thinking outside the box and checking and rechecking results. It also stressed the responsibility of scientists to always do their best work, which includes not publishing information they know is inaccurate. 

This interview has been edited for length and clarity.

Share your best worst mistake with us.

Submit your Story

Scientists have associated many neurodegenerative disorders including Alzheimer’s, Parkinson’s, Huntington’s, and prion diseases with protein misfolding and aggregation, where incorrectly folded proteins act as seeds converting properly folded proteins into the disease form.¹ This leads to insoluble protein aggregate formation. Previously, researchers inoculated animal models with samples derived from humans or animals with a suspected prion disease, such as Creutzfeldt-Jakob disease, bovine spongiform encephalopathy, or scrapie, to detect misfolded prion proteins and estimate their levels.² However, these bioassays were time-intensive, taking several months to complete, and were expensive. 

To overcome these limitations, scientists developed the real-time quaking-induced conversion (RT-QuIC) assay for both research and diagnostic use, where prion seeds taken from specimens induce protein aggregation inside a microplate’s well.³ Because of intermittent shaking and the incubation temperature, scientists can complete RT-QuIC assays in as short as 20–60 hours, while maintaining or exceeding the sensitivity of animal-based approaches. Additionally, researchers have also adapted this bioassay to study protein aggregation in Alzheimer’s and Parkinson’s diseases.

The scientists that originally developed the RT-QuIC assay employed BMG LABTECH’s FLUOstar® Omega microplate reader due to its ability to withstand the harsh and prolonged shaking cycles required for this experiment, while periodically quantifying the samples. Using this instrument, researchers can perform the bioassays continuously over multiple days, increasing their throughput and efficiency. Thanks to its innovative capabilities, scientists worldwide continue to employ this microplate reader for prion detection and aggregation assays of other misfolded proteins with neurological relevance, such as α-synuclein, amyloid-β, and tau.

Learn more about this microplate reader’s advanced features. 

What methods do you use to measure protein aggregation?

Submit your Answer