Nordic Life Science 1
A DVER T OR I A L PREVIOUSLY FOCUSED ON HIV resea
rch, the Infectious Disease Unit Research Department at Aarhus University Hospital is now heavily involved in research into COVID-19. To do this, as Dr. Mariane Høgsbjerg Schleimann, project manager for the department explains, they’re making use of The MACSQuant Tyto to bring their own considerable expertise in immunology to the fight. Tell us about your team! We’re made up of three individual but closely interlinked groups comprising around 40 people in total, all of whom are looking at infectious diseases and clinical medicine on a daily basis. Our HIV research focused on the discovery of neutralizing antibodies, a new therapeutic approach with great potential, and which we’re hoping will show similar promise in corona therapy. How does Miltenyi Biotec’s MACSQuant Tyto Cell Sorter fit in? Our intended workflow is to get plasma from recovered COVID-19 patients, incubate it with the virus, and measure the levels of neutralization. Good neutralizers will be tested against the 2002 SARS virus. After that, we’ll try to isolate B cells that can affect both viruses – the critical step for harvesting antibodies that can be used in treatment. The Tyto is extremely useful in this regard because we can use it on plasma cells, memory B cells, or whatever our targets will be. The safety advantages over traditional sorters attracted you the Tyto initially, but is sorting B cells from recovered patients dangerous? There’s no evidence it is, but we prefer to take no chances. We don’t have access to a cell sorter in BSL3 rooms (recommended for working with infectious samples and aerosol-producing lab equipment), so a regular droplet sorter is completely incapable of meeting our needs. The Tyto’s cartridge-based system makes it safe to move our samples around to BSL2, or even BSL1, because it’s fully closed. How does the neutralization assay work? First, we isolate antigen-specific B cells using the Tyto. Next, we perform single-cell DNA extraction to PCR amplify the gene sequences for the antibodies, which we then use to build and reproduce the ones we’re interested in and which we use to attempt to neutralize the virus. How many promising antibody candidates do you expect to identify in your approach? We’re planning to produce antibodies from 200 antigen-specific B cells per each group of five patients that we would further analyze in a 96-well– based high throughput screening assay. What benefits do you see from the MACSQuant Tyto? It requires no in-depth training. Once set up, a lab manager can be trained to use it easily and without flow cytometry experience, but being able to rely on closed, sterile containment is by far and away the major benefit. Sorting is what created a safety issue, and was the most critical part. The Tyto makes everything more practical as there no longer is a critical step. Can you imagine a COVID-19 research application where contamination risk would be a concern all the time? Anywhere in vitro or ex vivo cells have been infected and the interest is in the infected cells only. High concentrations of virus that would necessitate a BSL3 setting would absolutely benefit greatly from a Tyto. What have been your overall impressions of working with Miltenyi Biotec? They know us well, they understand our needs and make great recommendations. We’ve worked with them for years, and we love their products!