Member Info for Monkshood

It will be interesting to see how all of this pans out. Using magnetic beads can cut out centrifugation steps , but then you also have to give the affimers time to bind to the beads. What is the capacity of a bead relative to recovered virons with a precipitation step ?(personally I have found acetone ppn variable).
Cleaner samples can mean less time per sample on the MS (better signal to noise) and it should keep the MS front end cleaner.

The trace shown in the last Avacta presentation was the result of cutting up an affinity captured S1 protein into smaller pieces with a enzyme (trypsin) after it had been captured on a bead coated with Affimers. What is not clear is if this digestion step is required to get a unique diagnostic or if looking at the intact S1 was enough. There have been a couple of other pieces of evidence lately that using the intact protein is OK as a screen which is backed up by this proposed test. This is important as it makes things a lot faster and more reliable than doing a trypsin digest (although this will have its place in research).
What they are doing in this assay is just precipitating the proteins (including the Sar Cov2 virons) by just adding acetone. This is clearly cheaper and easier than using an affinity purification step (BAMS), you would end up with a less clean sample, but if it works - then does this matter?
Other aspects of the assay may well be similar - using UV to inactivate the virons , water as a mouth rinse (salt can interfere with the MS). Using reducing agents (beta mercaptoethanol or DTT ??) to break the cysteine cross linkages .

The positive is that they are clearly measuring intact S1 protein, so no trypsin digestion. But if an acetone precipitation works well enough why bother using BAMS?

Interesting find.
There is no indication when these were rejected. I think that the NHS stipulates 70% EtOH, where as at the start Crl was using 60% in their standard hand gel, so it could have been some early batches?

No idea, I can only assume that 'lets explore' is selling very well.
Maybe the idea to get them through covid this winter will turn out to be a significant move for the company and go on to make a larger contribution to earnings than the attractions?

Hopefully the 35p sp increase this week will have compensated....

I feel it deserves a bit more than being lost in a Humm thread...
As promised a November trading update.. Not sure why they released it at 11.00 though?
Since the end of June, they have accumulated 30M in cash, with a overall increase in net assets of 7M to 134M; on the downside the pledge book has reduced by 8M. Watch and jewellery sales have been strong.
They are keeping the shops open through lockdown this time. Instore viewing of watches/jewellery has stopped but they have extended the returns period for online. I can see sales in these areas being strong over Christmas so lets hope the lockdown does end in England on the 2nd. Note, they have a lot of shops in Scotland which will not be restricted in what they sell.
Their online portal for pawnbroking and loans is working now, so they are in a much better position than the first lockdown.

They have appointed someone to deal with the HTSTC loans issue. Given that revenue less impairment for these totalled £11m it should not be a major issue for them. I have been monitoring DebtCamel which is the ‘ go to’ place for complaints and advice about this (and other) debt issues, in over a year, I have only seen HAT mentioned once so it does seem to have the same issues as some of the other lenders.

Going forward, have now got 30M they can loan without going into debt (65M with their RCF).
Hopefully the update will bring a few more buyers in, increase the liquidity and lift the share price up (although it has been a good opportunity to pick some up at bargain prices)

Re -auditors , my partner used to work for one of the big four, it is not that uncommon for companies to switch, it is a bit like not switching your insurer , if you don't the price increases faster than perhaps it should..
It could just be that where things were tight in the first lockdown they looked at how they could save money and so put the audit out to tender.
I do not get the impression that they are bitter about being open, rather the converse., they flag it as a positive.

CEO - retires- he will be 70 next year! He is staying as a consultant for six months. Hardly a red flag.
It has been planned for a while, hence the recruitment of Chris Gillespie in the summer.

The auditor was out to tender and the outgoing Deloitte's say 'there are no circumstances in connection with its resignation which it considers need to be brought to the attention of the Company's members or creditors'.
Also the CFO (along with the new CEO) purchased shares at 267 in September, so I am not expecting him to be leaving anytime soon.

Nice bounce today; I assume that the market held off until index trackers had unwound their positions in SDL.
I have this as one of my core long term holdings now - time will tell if this proves to be a good decision but it has the potential to be a real global leader.

Interesting thanks. It amazes me how some of you find these things.
What it does illustrate, in common with all of the lab based methods, how much of the work is in the sample transfer at the start of the analysis. It may take one tech to 'run' 1k samples but it takes three to prep the samples.

Surely it is more to do with lockdown 2 than auditors?
I bought some at sub 200 today, I may not time the bottom on this but i am confident they are not about to go bust and will be a good mid/long term holding. The Mcap is now 81M , with no debt and net assets of over 126M(at the intrims).
They are classed as an essential service and have said they will not be closing this time. There may be some impact on Jewellery/watch sales but they have online and can use click and collect to keep these sales going.

This was to get rid of the mucin , not bind S1.
You could bind S1 as a clean up but then the buffer you would need to release it would stop it binding in the LFT. (or you could leave it bound and then bind a second Affimer with a reporter - but you then get a sandwich ELISA - which they already have!)
After a bit more reading the mucins self polymerise so they could be depleted in part, by filtration. It will be interesting to see what they do use pre LFT.

Strangely, this then links to the other aspect of Avacta, targeting cancers (although may need a different specificity).

Monoclonal antibodies, reactive with mucin glycoproteins have shown promise clinically for the diagnosis and classification of breast tumours, and they may also be used therapeutically for tumour targeting.

https://cancerres.aacrjournals.org/content/canres/48/8/2138.full.pdf

Perhaps they should coat the collection tubes with Affimers that bind mucin proteins?
Another product to market??

I just had a quick read, the viscosity comes from mucins. These are glycoproteins which is essentially what the S1 is, this makes it harder to have a prep method that separates them (pre LFT). I would think that it could also give problems with the HGPR type assays.
I can see why it is tricky getting good assays.
http://www.vivo.colostate.edu/hbooks/pathphys/topics/mucins.html

I don't think that they have any issues with mutation at the moment. I though that it was more about scaling production?
With regards saliva and LFT have a look at this -(it has been posted before I think)
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5469882/
And for a solution to get better samples....
https://4saliva.com/products/pure-sal/

Fard, I agree. but I would not want to overstate mutation being a major problem.
The initial screen took 4 weeks but to scale production and incorporate them in an assay, probably longer?
However, I doubt they would wait, they probably have already selected others. I think that they already have them for the nucleocore protein as well. As I said yesterday, there is a lot of sequence data now to see what is tightly conserved, so with careful selection the issue of mutation at the binding site can be minimised. A similar rational is used when selecting primer sequences for PCR.

Jaylarc, I had seen this but not found any further details. They clearly believe it is a feasible way.
I have not done much with glycoproteins, however, my concern is that although they say ' the chains of sugars are constant' my understanding is that the sugars and level of glycosylation can be variable and are dependant upon the host cell.

From the link below-
'A mixture of oligomannose- and complex-type glycans can be found at sites N61, N122, N603, N717, N801, and N1074 (Fig. 2). Of the 22 sites on the S protein, 8 contain substantial populations of oligomannose-type glycans, '

It is the one of the things that I would like to know about the BAMS assay, you could select a couple of tryptic peptides which are not glycosylated (this involves a slow sample work up), but if you look at the intact S1 protein (fast), then you will have multiple glycosylation states (therefore different m/z's), it may be that in practice you just get a broad peak which is still unique enough to be diagnostic . It is why I asked for details - but he never answered this part of the question.

https://science.sciencemag.org/content/369/6501/330

The only way an Affimer could directly detect a mutation is if it specifically bound to the epitope generated by the new AA (and not the old), or by loss of function i.e. it bound the original but not the new one.
You could see some mutations by MS following affinity purification by an Affimer - but that is a different thing.
I have only seen reference to AS saying that binding of the Affimers they are using is not affected by the G614D mutation.
Or have I missed something ?
Forever, no idea, I think most are using mAbs so it would be a similar issue, tests using pAbs (polyclonal antibodies)bind more sites so it would reduce the mutation problem but they usually have more problems with lack of specificity.
The virus is being sequenced at multiple sites all of the time. Changes would be quickly flagged, they could potentially affect PCR as well (if the mutation was at the terminus of the primer site) so it is not entirely unique to protein detection based systems.