Polycarbonate Ultracentrifuge Tube Re-Use in Proteomic Analyses of Extracellular Vesicles

We study cardiovascular diseases, specifically the role of extracellular vesicles in the pathological calcification of cardiovascular tissues, such as valves and blood vessels. Researchers in our group have recently defined methods to extract tissue and trapped extracellular vesicles from fibro calcific cardiovascular tissue to determine their proteomic profile. Our group has demonstrated that these extracellular vesicles contain important protein cargo for initiating calcification in diseased tissues.

The specialized tubes used in the ultracentrifugation step of extracellular vesicle isolation can be cleaned and sterilized for reuse. However, proteomic analysis requires special consideration as any contamination could greatly affect downstream results. Here, we validate a protocol for reusing consumables, specifically polycarbonate ultracentrifuge tubes for extracellular vesicle isolation that is suitable for downstream, low input proteomics applications.

Our protocol is simple and effective and it will allow researchers to reduce both the consumable costs and the waste typically associated with these isolation protocols. To begin, gather used polycarbonate tubes and discard the previous sample thoroughly. To prevent drying of the sample to the side of the tube, immediately submerge the ultracentrifuge tubes in tap water.

Then incubate the tube in 0.1%sodium dodecyl sulfate, or SDS, for 10 minutes. Decant most, but not all of the SDS solution from the tube. And using a cotton swab thoroughly wipe the area of the tube that had the extracellular vesicle pellet.

To rinse, fill the tube with hot tap water and decant completely. After three rinses using a spray bottle, spray demineralized water into the tube followed by 70%ethanol. Place the tube upside down to dry completely and transfer it to a clean jar for reuse.