Cell Sorting is the process where we trigger a sorting mechanism based on the fluorescent intensity of a cell. Importantly this gives us the flexibility to gate based on either internal markers such as GFP, autofluorescence, or FoxP3 or multiple externally tagged specific markers. This allows us to separate out complex cell types of interest from the bulk population. This is an incredibly powerful technique where we can significantly increase the specificity of the cell type that we’re isolating and the types of signals that we can use for that isolation.
Who Should be Most Interested in Cell Sorting?
The people that are likely most interested in cell sorting in the clinical arena would be translational scientists developing a CAR therapy (CAR-T, CAR-NK, CAR-M, or CAR-Treg, which we will collectively call CAR-X). At KCAS, we have developed very special techniques where we can quickly sort out these extremely unique cells from patient cells as part of a clinical trial. We can sort as few as 1,000 cells in under 2 hours directly into an extremely small volume of lysis buffer- a workflow developed specifically to avoid biasing signals in downstream RNA analysis workflows. This is extremely powerful for understanding exactly what lineage the cells are from, how their modifications are expressed, and what type of RNA is still being expressed in the cell. This carefully orchestrated workflow is an excellent example of the ways that we’re using our sorter in the bioanalytical sphere.
The other primary group of people that are interested in cell sorting are researchers in the discovery space who are tagging cells with different fluorescent proteins and using that fluorescent protein expression for downstream isolation. This can be an extremely laborious process where you need to manually pick out each cell and then put it somewhere else. Instead of doing this manually over time, we can carry out the same process with up to one hundred million cells an hour – greatly accelerating the pace of their research and ideally getting the highest expressing cells and making sure they’re getting a representative view of those cells (as opposed to what one can do with a microscope or with a few plates, for instance).
Solving the Next Generation of Technical Challenges for Cell Therapies with Cell-Sorting
For CAR-X developers, the biggest headache that we hear about is the difficulty of finding a CRO or clinical site that can isolate these CAR cells in a useful way. Most often these providers can isolate PBMCs or in some cases use magnetic enrichment to get bulk cell populations, which doesn’t get developers the specificity they really need for their custom antigens. At this point, their options are either to attempt to conduct this in house, find a university and go through their infamously complicated contracting process (and potentially lose the privacy of their data) or… work with us.
The availability of these cell-sorting services overall in the industry are limited, and therefore something we really pride ourselves on. It is something we have been doing for the entirety of our PA facility’s history – nearly a dozen years at this point.
The Right Equipment for Discovery Research as well as Clinical Trials
At KCAS, we have recently upgraded to the BD Biosciences FACSymphony™ S6 6-way Cell Sorter instrument. This is the latest and greatest from the BD family. It can analyze up to 30 colors simultaneously and has high-power lasers to make sure that we can dig down into even very, very dim signals. As its name implies, it allows up to 6-way sorting, so if we need six bulk populations, we can do that and get a lot more per mill of input volume we are sending in.
If we sort single cells into each well of a 96 well plate, we can actually trace the cell in the plate back to the event in the flow plot thanks to our indexing software. Especially for clone picking and discovery work, this can help researchers relate downstream analysis back to how intense their fluorescent expression was during the sort. This is an incredibly powerful upgrade over our previous system and is truly an industry-leading capability in this space.
In addition to these unique workflows and cutting-edge technology, our value proposition for any project always includes our people.
Our people have the most expertise in doing this. They’ve done this with fluorescent proteins and other very simple sorts. They’ve done this with CAR proteins and very specific sorts for clinical studies. We’ve done this very special technique that allows us to get RNA signatures afterwards with minimal disruption to the cells during this sorting process. We can work in lockstep with our internal LBA, Mass-Spec, or PCR groups to rapidly develop a full processing pipeline under a single contract, or we can engage with external groups if we need to leverage other sponsor relationships.
Our equipment, experience, and expertise set us apart and give us the ability to identify and overcome challenges that might come with a given program. Our sponsors can trust that they have access to the latest and greatest options that are available with modern technology, and that with our on-going global growth and investments in both people and technology we will continue to put them on the forefront of what’s possible.
If you have any questions about these services or any others offered by KCAS, please use the form below. Thank you!