Protein Related to ALS Onset Used to Develop New Method of Antibody Validation for Use in Research

Protein Related to ALS Onset Used to Develop New Method of Antibody Validation for Use in Research

Scientists have developed a new method to validate antibodies for research, demonstrating its effectiveness by using the protein encoded by the C9ORF72 gene, which, when mutated, may cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), a study reports.

They also urged its implementation in labs around the world to avoid data inconsistency and lack of reproducibility in scientific research.

The study, “Implementation of an antibody characterization procedure and application to the major ALS/FTD disease gene C9ORF72,” was published in the journal eLife.

Antibodies are molecules designed to recognize specific regions of a protein. They serve many different purposes in biomedical research, including allowing investigators to study protein localization within a tissue or cell, as well as to measure its levels.

“Antibodies are vital tools in the biomedical research arsenal and in keeping with their importance there are over a million unique antibodies that are commercially available,” the researchers wrote.

However, little attention has been paid to antibody characterization or to the implementation of standardized procedures to ensure the quality of this process.

“As a result, most products are generated and then sold/distributed with only rudimentary and non-quantitative supporting data, and as a result there are serious flaws in the reliability of many of the available reagents,” the authors wrote.

This has raised concerns among the scientific community regarding the validity of findings reported in studies using non-validated commercial antibodies, and called for the development and implementation of a new method that would allow investigators to assess the quality of a specific antibody of interest before using it in their experiments.

In this study, researchers at The Neuro (Montreal Neurological Institute and Hospital) and collaborators described a new method to validate antibodies, using the protein encoded by the C9ORF72 gene as a test case.

“C9ORF72 provides an excellent protein on which to develop an antibody characterization process because although the protein is of relatively low abundance, there are many commercially-available antibodies,” they said. “The process we outline can be applied to any protein target and here it led us to the recognition of problems with the C9ORF72 literature.”

After testing 16 commercial antibodies marketed as being specific for C9ORF72, the team discovered that only one of these antibodies was able to detect the C9ORF72 protein in immunofluorescence, a technique that allows researchers to see under a microscope where in a cell or a tissue a protein labeled by a specific antibody is present.

In other tests, the team found that only two other antibodies seemed to specifically recognize the C9ORF72 protein.

In addition, after examining the literature, they found that none of the antibodies that passed their tests had been used in any study, while several of those that failed to meet their validation criteria had been used in many studies.

These findings question the validity of observations made in previous studies that used antibodies that were either unable to recognize the C9ORF72 protein altogether or detected additional unspecific protein. They also highlighted the importance of establishing a standardized antibody validation protocol that can be adopted by labs worldwide.

“As we worked on our C9ORF72 paper, it became less about one gene and more about a template other labs can use to validate antibodies,” Peter McPherson, PhD, corresponding author of the study, said in a press release.

“The procedures we use are not revolutionary, and in fact this makes our approach widely applicable to any laboratory skilled in the art, yet to my knowledge this is one of the first papers to describe a streamlined process for antibody validation. A large part of the reproducibility crisis is because of poor antibody validation. We owe it to funders and patients to do better,” added McPherson, who is a professor of neurology and neurosurgery and anatomy and cell biology at The Neuro.

Joana is currently completing her PhD in Biomedicine and Clinical Research at Universidade de Lisboa. She also holds a BSc in Biology and an MSc in Evolutionary and Developmental Biology from Universidade de Lisboa. Her work has been focused on the impact of non-canonical Wnt signaling in the collective behavior of endothelial cells — cells that make up the lining of blood vessels — found in the umbilical cord of newborns.
Total Posts: 279
Margarida graduated with a BS in Health Sciences from the University of Lisbon and a MSc in Biotechnology from Instituto Superior Técnico (IST-UL). She worked as a molecular biologist research associate at a Cambridge UK-based biotech company that discovers and develops therapeutic, fully human monoclonal antibodies.
×
Joana is currently completing her PhD in Biomedicine and Clinical Research at Universidade de Lisboa. She also holds a BSc in Biology and an MSc in Evolutionary and Developmental Biology from Universidade de Lisboa. Her work has been focused on the impact of non-canonical Wnt signaling in the collective behavior of endothelial cells — cells that make up the lining of blood vessels — found in the umbilical cord of newborns.
Latest Posts
  • neuronal degeneration, SARM1, ALS
  • ProMIS
  • AP-101
  • CuATSM Phase 2/3 trial
Average Rating
4 out of 5 stars. 3 votes.
My Rating:

4 comments

  1. Bobbie Dreher says:

    I believe you are on a better track for this. Carrying this gene in my family have it myself with symptoms now. I am doing vitamins and CBD and Thc in very small amounts only. On a by pap machine at night. I feel Als strips us our vitamins certain ones With my potassium was first then Vit D. I take 1000 vit C daily, I will continue this and hope it helps others. When I do this it keeps my saliva dry, I don’t cry, I am doing well mobile. But I do this every 6 hours 3 times a day most times. It has stopped my facilitations and cramping of feet and legs. It stops the spasticicty, I sleep well and I can eat and swallow. I have been on my by pap for going into the 3rd year and have asthma. my oxygen is 99,98. And upping my cbd as was having trouble sleeping I had 6 nights no events on my by pap. never happened before.

  2. Debbie Chalk says:

    My husband died of ALS 2 years ago today. It would be wonderful to find a cause and a cure. It is a horrendous disease, to see your loved one who was once so very strong have all his muscles waste away. He was only 64 years old.

  3. Mariam Donerian says:

    I find the first comment very useful.my boyfriend had also, just died in August. He liked vitamin C too. Use to buy those tablets. But I use to buy those little oranges, peel them and he would eat the slices one by one.it helped restore his voice alot too.but he was no fan of ventilators, c-pap machines, cough machines, and the like. He would only use his Albuterol once in a while. That restored his voice also. He was supposed to use that 4 times daily. We were on a race against time, and we did the best we could. His progression was fast,and he wanted to live on his own terms.

Leave a Comment

Your email address will not be published. Required fields are marked *