Opentrons Customer Interview | Oscar Swindley, University of Sheffield

Oscar Swindley works on monoclonal antibody folding and assembly at the University of Sheffield. He titers cell cultures using an Opentrons OT-2 pipetting robot.

The University of Sheffield is one of the most influential research universities in the UK. It has a long history and ranks 95th in the 2022 QS World University Rankings. . Oscar Swindley is Chemical and Biological Engineering at the University of Sheffield Ph.D., engaged in monoclonal antibody folding and assembly. He uses Opentrons OT-2 pipetting robots for titration assays for cell culture development. Today he will share with you his experience using OT-2.

Oscar Swindley is a PhD student in the Department of Chemical and Bioengineering at the University of Sheffield, where he is part of the team developing the synthetic biology platform. The laboratory purchased its first Opentrons OT-2 in August 2019 for cell culture development. For titer determination automation, Oscar is an in-depth user responsible for operating OT-2.

Opentrons: Please introduce your research background?

Oscar Swindley: My research interests are in the genetic and chemical manipulation of monoclonal antibody folding and assembly processes, as well as the optimization and integration of automated procedures. We use OT-2 to perform a large number of cell culture procedures in our laboratory. I just completed our first large screening project where all cell culture, transfection, sample collection and titer determination were performed on the OT-2.

Opentrons: How would you describe a day in the lab?

OS: Recently I was working with the results of the first large-scale cell culture screen done on the OT-2. I have been using the OT-2 for titration. OT-2 supports and simplifies the entire process. Apart from this, my regular day-to-day work includes routine cell culture in shake flasks and well plates, as well as other experiments in the molecular biology laboratory.

Opentrons: Why did you choose OT-2?

OS: The purpose of introducing OT-2 in our laboratory is to improve the reproducibility of our platform. We use 24-well and 96-well plates for high-throughput screening in our laboratory. The main difficulty we faced was with transfection, because when we resuspend the cells at very high densities, the solution becomes viscous and the cells settle quickly. We need to accurately distribute the sample across the well plate so that the same number of cells is grown in each well. This is a highly repetitive process and deals with very small volumes of liquid. The biggest problem we faced was how to always keep the liquid well mixed and successfully seed the cells while the cells would gradually settle over time.

Opentrons: What exactly do you use the OT-2 for, and how does it fit into your workflow?

OS: We use the OT-2 to set up and run our 96-well electrofection platform, sample from multiple culture plate formats, and perform plate-based assays. Specifically for our electrofection platform, OT-2 combines DNA, buffer, and cells and then distributes the mixture into the electrofection plate. After transfection, OT-2 is diluted, mixed, and supplied with nutrients to the cells, which are then seeded into 24-well culture plates. After 3 days we will assay the cultures using the robot. This includes mixing and sampling from culture plates for various downstream assays. Finally, the OT-2 is capable of loading samples onto glass slides for use in our high-throughput cell counters and harvesting supernatants for titer analysis. In a recent set of screening experiments, OT-2 helped us perform 22 electrofection plates and associated sampling. The OT-2 can run a titer analysis in just 20 minutes, which is great. I can keep the bot running all day long.

Opentrons: What was it like to start experimenting with your OT-2?

OS: It took me about 2 months to write my first protocol script. But once the script is complete and all the information is in the script, I can quickly move on to more complex experiments. I uploaded all my scripts to my GitHub page. If you are interested, everyone can check it out. .

Opentrons: Did you use other laboratory automation equipment before using the OT-2?

OS: No. I have used BioMek before and have SOPs set up. To get better at programming the OT-2, I learned Python from August to November.

Opentrons: Do you have any other experiences about OT-2 that you would like to share with everyone?

OS: Overall this is a good device, but it does not have a safety device, which means it cannot recognize obstacles or realize if the suction head has fallen. The problem we have now is being able to track failures when they occur. Another difficulty was that the device was not very compatible with 24-well plates, so I had to find a way to cheat it by removing tips from every other row. But if you know how to program a robot, you can always find a solution. Overall, I think it's great for research, and you can tailor the protocol to your needs in each experiment.