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New Metal conjugation kits from expedeon

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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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ShreyaSharma

Participant

Posts: 1

Joined: Thu May 23, 2019 12:20 pm

Post Wed Oct 30, 2019 1:20 pm

New Metal conjugation kits from expedeon

Hello everyone

Has anyone tested Lightning-Link® Metal Labeling Kits from Expedeon
I understand that this kit uses metal chelating polymer that cross-links through primary amines. While the Fluidigm uses different labelling chemistry.
Are Expedeon kits good for labelling plus how stable is the conjugation upon storage? also are there any chances of this chemistry not being compatible with other labelled antibodies in the Panel.

Shreya
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mleipold

Guru

Posts: 2086

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Oct 30, 2019 6:51 pm

Re: New Metal conjugation kits from expedeon

Hi Shreya,

I have not tried the Expedeon kits.

The main issue: I cannot get a straight answer out of them on how many chelators per "warhead". Based on their description, I *think* it's one chelator per warhead, similar to the ITCB-EDTA or mDOTA.

I also would point out their signal intensity here: https://www.novusbio.com/products/light ... _m139-0100 There, it appears CD45 is only giving a 139La intensity of ~1e2 Dual, about 1/10th of CD45-Pr141 (https://www.fluidigm.com/reagents/prote ... --100tests) or even CD45-89Y (https://www.fluidigm.com/reagents/prote ... --100tests) from Fluidigm.

If that's the case, then I'm not sure what markers would be easily conjugated and still have acceptable signal.


I also spoke recently with Bio-techne/Novus Biological who are re-sellers, and they couldn't tell me either.


There's also a cost issue: 1-rxn kit for 100ug labeling is listed as $749, which is more than a 4 rxn X8/DN3 kit from Fluidigm.


Mike
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MCOlivier

Contributor

Posts: 31

Joined: Mon Oct 05, 2015 9:48 am

Location: European Genomic Institute for Diabetes, Universitity of Lille, Institut Pasteur de Lille, France

Post Thu Oct 31, 2019 9:35 am

Re: New Metal conjugation kits from expedeon

Hi all, and thanks to launch this subject !

I bought 1 139La kit and coupled my anti CD8 this week. I'll test it in a few days and keep you awared of the signal I get.
Considering the cost issue, I really hope it will be satisfying !!!
Regards ;-)
Olivier
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DMcDonald

Participant

Posts: 11

Joined: Thu Nov 26, 2015 10:14 am

Post Thu Oct 31, 2019 12:29 pm

Re: New Metal conjugation kits from expedeon

Hi All,

I did some limited testing of these about 18 months ago during a beta-test. I conjugated a huCD45 to 159Tb and had signal >10e3 duals. I haven't retested this Ab though recently which would be a nice to see the stability. Hopefully I'll have the chance to test some more of the kits now they're on the market as their product format for 10 ug reactions make them very amenable to many of the antibodies we get for IMC projects where you struggle to get 100 ug at an affordable price.

The two major benefits were recovery and time. As there are no spin steps then you can't really lose any Ab and it takes minutes not hours to perform a conjugation. Despite the superior recovery, they do have their pricing wrong though it should be brought more in line with Fluidigms pricing.

Looking forward to hearing how you get on as some healthy competition has been a long time coming in the field.

Best wishes,
David
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Azurelexa

Participant

Posts: 1

Joined: Fri Nov 22, 2013 2:35 pm

Post Fri Nov 01, 2019 4:15 pm

Re: New Metal conjugation kits from expedeon

Hi All,

we have recently tested two 10 ug reaction kits from expedeon, 139La and 169Tm. Both kits were tested in parallel with an in-house anti-human CD8 clone (GN11/134D7).
The protocol was very quick and simple. We tested both conjugates on some frozen PBMC and compared them to a Maxpar-conjugated CD8 antibody. Essentially, the 169Tm Kit worked very well (see attached pictures) and generated the expected staining pattern on PBMC, which was comparable to the Maxpar conjugate. On the other hand, the 139La Kit did not produce any specific signal, even when used at very high concentrations.
I also attach a screenshot from the acquisition of the 169Tm CD8 conjugate, where you can see that the conjugate is a bit streaky, which probably is due to some unbound 169Tm in the antibody solution. But I think this is a tolerable level.
As we have recently conjugated the antibodies and performed the test only once, I cannot tell you anything about the stability of the conjugates.

https://ln2.sync.com/dl/3f6c80cf0/pzyv9 ... 2-dgg98xdg
https://ln2.sync.com/dl/500560a30/pzspd ... 2-rvsbmfnn

Best,

Axel
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DMcDonald

Participant

Posts: 11

Joined: Thu Nov 26, 2015 10:14 am

Post Mon Nov 04, 2019 2:15 pm

Re: New Metal conjugation kits from expedeon

Hi All,

Just a quick update, I did a quick stain with the conjugation I did early last year alongside a similar CD45 Ab from fluidigm of a similar age. The Fluidigm one looked fine still but the lightning link Ab no-longer showed any cell-associated signal but rather some decent intensity irridium negative events. Presumably these were large antibody aggregates. I'd stress this was an early version of the kit so might not translate to the performance of the commercial release but it's worth keeping a close eye on their longitudinal performance in case this problem persists.

Best wishes,

David
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DouglasLinn

Participant

Posts: 2

Joined: Wed Nov 06, 2019 9:07 pm

Post Fri Dec 06, 2019 3:48 pm

Re: New Metal conjugation kits from expedeon

David,
How months would you say it was between the testing of the Lighting conjugated antibody(s)? Potential decreased shelf life is a little disappointing
-Doug

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