Validated antibodies for ALS research: the ALS-RAP project

We are part of ALS-RAP, the ALS (Amyotrophic Lateral Sclerosis) Reproducible Antibody Platform. We’re addressing one of the big conundrums to biomedical research: the lack, or the poor quality, of many antibodies used for research ranging from cell biology to histopathology.

ALS, also called MND (motor neurone disease), is a group of diseases that cause degeneration of motor neurones. The symptoms vary, but the disease is debilitating, life-threatening and currently incurable. Around 5,000 people in the UK, and 12,000 people in the US, are currently affected. There are charities in several countries devoted to helping patients and supporting research, including the ALS Association (US), MNDA association (UK) and the ALS Society of Canada; they all have very informative web sites. These three charities have joined forces to fund the ALS-RAP project

Up to 10% of cases are hereditary, and the genetic changes have been mapped for many of these cases. Mutations of C9ORF72 and SOD1, TDP-43 and FUS are most common, but there are dozens of other genes that are mutated less frequently. Studying the ALS-related genes and their roles in healthy nerve function and in disease are keys to understanding the biology and pathology and, eventually, finding cures. To do that, we need high-quality, validated reagents; in many cases, these do not exist (or we have no way of knowing, because we haven’t tested them properly!)

So here’s our plan. We have set up a collaboration with Susanne Gräslund at the Karolinska Institute in Stockholm and with Tom Durcan and Peter McPherson at the Montreal Neurological Institute (McGill University).

First, we consulted our advisory board to build an initial target list; this is mostly based on genetic data, nicely summarized here.

The Canadian groups are establishing a robust validation process: basically, using cells that express “native” levels of the target protein, and matched CRISPR knockout cells, to probe the sensitivity and specificity of antibodies. They have started to probe commercial antibodies

The Stockholm team has libraries of single-chain antibody variable domains (scFv) in bacteriophage; they will screen the library to isolate and characterize antibodies to each target protein. We also expect contributions of new antibodies from commercial collaborators, who will (of course) put these on their catalogues.

The Oxford team (that’s Tracy Keates and me) are expressing and purifying the target proteins as biotinylated, well-folded proteins or protein domains, to be used as antigens for antibody screening.

You can see the details in our web page: , including the target list, target status, plasmids and protocols, validation data (soon) etc.

In the next blog, I’ll tell more about our cloning campaign to generate the antigens.

Leave a Reply

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