Welcome to the SGC-Karolinska side of ALS-RAP: the scientists, the screening, and the antibody selections

Hello fellow scientists and open lab notebook readers!

I’m Carolyn Marks and I’m working in the ALS-RAP partnership with Susanne Gräslund, who leads the Recombinant Antibodies Group at SGC-Karolinska in Stockholm, Sweden. SGC-Karolinska has had a long-standing collaboration with Helena Persson Lotsholm, Director of the Human Antibody Therapeutics Facility within the Drug Discovery and Development Platform at Science for Life Laboratory in Stockholm. I am fortunate to work with and learn from several experts to generate the highest quality antibodies for the ALS-Reproducible Antibody Platform. I joined the ALS-RAP team in September of last year and from here on out I’ll be sharing live updates of our work with this community.

Perhaps I should tell you a little bit about me, what we do at SGC-Karolinska, and how I got here. I’m a wet lab kind of gal and a neuroscientist at heart with a lot of years of experience under my belt. The methods that I use here at SGC-Karolinska for the ALS-RAP are broad and range from phage display technology to cloning to high-throughput screening to patient-derived cell-based assays. Actually, many of the methods that I’m using now to generate and validate recombinant antibodies for the ALS-RAP, I used for many years to study neurodegenerative diseases, including ALS.

I´m originally from Texas where my love of neurophysiology began, and where I was initially trained as an electrophysiologist. From there, I deferred medical school and took an internship at NINDS at the NIH to have a break before med school began. My 1-year internship turned into two, and I quickly understood why everyone at NIH kept telling me “Carolyn, you reek of a PhD!” I soon found the NIH-Karolinska Graduate Partnerships Program and hopped on a plane. That’s how the girl from Texas ended up in Sweden. I received my PhD in 2012 from the NIH-Karolinska Collaborative Doctoral Program in Neuroscience; and I recently completed my Post-Doc, also at Karolinska, characterizing the ubiquitin proteasome system as therapeutic targets in neurodegenerative diseases.

How did I find my way to the SGC, you might wonder? In my research over the years, I have experienced first-hand the need for reliable high-quality antibodies – and the lack thereof, as well. I hate that we as scientists are limited by the tools that companies provide. These limitations are crippling our research. Sadly, the ones who suffer the most are the patients. This was my motivation for joining SGC-Karolinska and particularly this project. I wanted to make better tools for the research community. Consequently, I believe that better research tools will yield better science and hopefully better therapeutics for the patients who need it. That’s why I’m here.

How does the ALS-RAP work? Our team at Karolinska is only 1 of many. We work together primarily with scientists at Oxford and in Montreal. The ALS-RAP was created by three major ALS charities from Canada, the UK, and the US. The long-term goal of this partnership is to provide the ALS research community with reproducible, thoroughly validated antibodies for selected ALS-related proteins. This includes testing existing commercial antibodies as well as generating new ones. The work is done in three different labs, as recently outlined by my fellow ALS-RAP partner and open-lab-notebooker, Opher Gileadi.

In the Stockholm team, we use phage display technology to create recombinant antibodies. Don´t know what phage display is? The inventors of this technique were actually awarded the Nobel Prize in Chemistry in 2018. While this technology has many uses, we use phage display to generate recombinant antibodies. How does it work you might wonder? When we receive antigens of our ALS proteins from our friends in Oxford, we then use the phage display technique to select out antibodies that binds these antigens. We use extensive libraries of human antibody candidates where high-quality binders can be obtained with the phage display method. We then perform several rounds of screening and selection to ensure that we get the best possible antibodies. We then evaluate their specificity and affinity and finally we validate the antibodies using immunoprecipitation followed by mass spectrometry. It sounds simple, but trust me it’s a thorough process! Last but not least, we will release the sequence of the best antibody for each target on our website and make it available to anyone who wants to use it. Our collaborators in Montreal are creating CRISPR-knockouts of these targets in both standard cell lines and iPSCs for further validation and application specificity.

We have a really exciting target list, built by the help of our ALS-RAP advisory board. Check it out and see if your favorite protein or that much needed antibody is on our list. If not, click here and request it. So far, we have generated and selected our first set of antibodies for 7 of these targets. Now onto primary and secondary validation.

Stay tuned to find out who the lucky 7 are…

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