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Magnetic beads selection - ?interference

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It is fine to promote your company's reagents. Just make sure they are relevant to CyTOF, and do so in moderation and style :-)
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Ondrej

Participant

Posts: 5

Joined: Fri Feb 16, 2018 12:27 pm

Post Mon Sep 17, 2018 4:13 pm

Magnetic beads selection - ?interference

Hi everyone,

we are about to run a large collaborative project where I will be given positively selected human lymphocytes for CyTOF. I am worried that the magnetic beads might interfere with other channels. I see that people advice to use negative selection if possible, but that is unfortunately not an option for me.

a) Do you know if Miltenyi magnetic beads used in selection kits cause problems? If yes, is this interference with one particular channel or multiple channels?

b) Do you know of any other company which might be selling CyTOF friendly selection kits (based on magnetic beads)?

c) Some people are suggesting to strip off the beads from the cells before running CyTOF. Do you have any protocols how this is done? Does such treatment affect other surface markers?

Many thanks for your comments to any of my questions!

Best regards,

Ondrej
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Mon Sep 17, 2018 4:29 pm

Re: Magnetic beads selection - ?interference

Hi Ondrej,

Regarding part c: Miltenyi has their MultiSort product line, which cleaves the bead off the enrichment reagent (eg, biotin: 130-091-256) to allow you to collect your enriched cells. The anti-biotin would still be adhered to your cells, but the beads would be left behind.

I have a couple labmates here at Stanford HIMC who have run these enriched cells without substantial background. They didn't mention anything about other markers being affected, but of course you'd have to try it in your system to be sure.


Mike
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tomaskalina

Participant

Posts: 10

Joined: Fri Dec 20, 2013 9:19 am

Post Mon Sep 17, 2018 5:18 pm

Re: Magnetic beads selection - ?interference

Hi Ondrej,

you can consider non-magnetic positive selection. There is automated system (FABian) for column based positive selection using Fab-streptamers by IBA.
https://www.iba-lifesciences.com/automa ... -shop.html

The don't have a huge marker list, but CD19, CD3, CD4, CD8, CD28, CD14 and CD81 is covered.
You can process the cells with the sequence of enrichment steps if needed.

Best regards, Tomas
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cguidos

Contributor

Posts: 28

Joined: Tue Nov 18, 2014 3:10 am

Post Mon Sep 17, 2018 6:07 pm

Re: Magnetic beads selection - ?interference

Hi Ondrej

We have run murine cells positively enriched for CD45+ cells using the Miltenyi beads and Auto-macs. Did not have any problems!

Cindy
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Ondrej

Participant

Posts: 5

Joined: Fri Feb 16, 2018 12:27 pm

Post Tue Sep 18, 2018 5:24 pm

Re: Magnetic beads selection - ?interference

Hi guys,

many thanks for your comments on how to get rid of the possible magnetic beads interference. I have been searching for any systems offering the beads removal after the selection and found this new kit from Miltenyi which seems pretty much suited for my purpose. It not only gets rid of the beads but also uncovers the epitope which would be normally covered by the "old-fashioned" antibody-bead conjugate.

Here is the link if anyone is interested as well:

https://www.miltenyibiotec.com/GB-en/pr ... 30-117-034

Does anyone have a good experience with this new kit?

Best wishes,

Ondrej

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