Literature/202301141333 comparing simoa and sp-iris

First published:

Last Edited:

Number of edits:

Simoa and SP-IRIS are used and compared in their abilities to detect extracellular vesicles. Both are digital methods (see: Analog versus digital detection of molecules). And they can be a crucial tool to detect and quantify Low abundance EVs in bio-fluid derived samples.

Using the SiMoA platform

The SiMoA platform was from Quanterix, and they used the Homebrew Kit (I assume a kit that allows custom bead functionalization). The concentrations of beads used was $2.8\times 10^9 \,\textrm{pcles}/\textrm{ml}.$ Sample, beads, and detector are incubated for 30min. Samples and then washed (automatically) and incubated again for 10minutes.

Using the SP-IRIS platform

The NanoView Biosciences instrument was used, with custom silicon chips. Chips were incubated for 2 hours, and then placed on a shaker for 3min. Fluorescent antibodies were added and the chips were incubated for another hour.

SiMoA results

images/Pasted image 20230114154800.png

images/Pasted image 20230114154816.png

SP-IRIS results

images/Pasted image 20230114154715.png

images/Pasted image 20230114154744.png

Comparison on same cell-samples

images/Pasted image 20230114155207.png

In the figure above, the relative expression of some tetraspanins for different cell lines is measured. Then, SP-IRIS and Simoa are used to measure those same proteins. The limit of detection of SP-IRIS is worse than that of SiMoA for low abundance specimens (CD63 in HEK cells, CD81 in HeLa cells, for examples).

It is interesting that the 'background' signal changes depending with the tetraspanin targetted. For example, in CD9, the 'Fluorescence Particle Counts' of SP-IRIS goes down to ~10, while for CD81 is always above 100. On the other hand, the SiMoA instrument goes from 0.001 in CD9 and CD81 up to 0.01 in the case of CD63.


Comment

Share your thoughts on this note
Aquiles Carattino
Aquiles Carattino
This note you are reading is part of my digital garden. Follow the links to learn more, and remember that these notes evolve over time. After all, this website is not a blog.
© 2021 Aquiles Carattino
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License
Privacy Policy