Covid I'm skeptical about. My lab requested inactivated covid for me to validate my assay on and we ate silence from them. We never had trouble getting innactivated Rabies particles from them for another thing we were working on.
That's good to know. Might I ask if you know what machines do you use for validating the assay and what corporation are they from? I have a strong suspicion against Illumina though I don't have much to go off on.
I'll think about it. It really hurts to see mindless bias masquearding as critical thought here. People are supposed to be here because they are BETTER than the left, not just the same retards but with a different dogma they blindly follow.
I agree, I do not like the mystical conspiracies you'll sometimes see here either.
I may be wrong on some things and I want to know them so I can learn and progress towards truth.
I am hesitant to detail the assay I would be using the particles as validation for as it would likely be enough to out me and my lab and living where I live has instilled in me an odd mix of caution for precisely when and where and how I stick my neck out.
In general terms, drug discovery requires a degree of throughput that can't be accomplished by doing things the 'best' way. One can't for example, infect tens of thousands of monkeys with SIV to test individual drugs against - ethically you've gone from a grey area to something utterly reprehensible. Doing so would also take a gargantuan amount of time and cost astronomical amounts.
It's better to do things in cell culture. Better throughput, even though as a collection of cells rather than a living breathing immune system or integrated organs it might not be quite as representative. Do the results in cell culture transfer to animal? Often not.
But even cell culture is low throughput, so in reality the only way to have a shot is to design assays which approximate the functions of relevance in the cells that are representative of the larger organism to test agaisnt the mechanism of action you're trying to develop against - for example, merely replicating the receptor binding that is prelude to entry, or the binding regions of a capsif they rely on for assembly.
But how do you know your assay which so removed from the integrated whole is actually representative? Well, you validate it. And for many such examples, trialing against actual legitimate viral particles is enough. If the virion binds the same as your stand in, and shows similar reactivity to some of your early drug candidates, you can have enough confidence (never needs to be 100%) that's it's then worth the time to roll the dice running that assy hundreds of times on tens of thousands of unique compounds to find one or two worth doing more intensie looks on...
...In cell culture. And if that works, in Animal trials. So as you could imagine, I am happy to receive other irradiation particles from the CDC, and a little perterbed when I don't know if my generic SARS1 coronavirus assay is sufficiently representative to SARS2 to be worth the time.
Dude I'm totally good with it, I'm at a university and I have some (albeit very very small) experience with what you're doing.
I'm not as interested in the assay but the equipment (or really the manufacturer) that measures everything.
(Correct me if I'm wrong) an assay is a substance that you get that is analyzed for
having our Covid-19 virus.
Throughput is the rate of production or the rate at which something is processed, so a cell culture gives little data to process but if you design the cell culture (collection of cells) around the cells the Covid-19 virus is supposed to drastically infect and alter, and if the assay (which is supposed to have the virus) binds to the cells the same way Covid-19 virus does.
If the virus (idk difference between virus and viron) binds the same way, you revalidate it with other cells and once you've sufficiently done that, you have the confidence that this assay is worth the time to run hundreds of times on tens of thousands of samples/compounds to see maybe one or 2 compounds that react differently (maybe for vaccine or interesting study).
So as you could imagine, I am happy to receive other irradiation particles from the CDC
What are irradiation particles?
Also it looks like you cut off towards the end, please tell me more when you get the chance! So your generic SARS1 coronavirus didn't represent SARS2? That's really weird, they should be practically the same thing with a better binding mechanism for SARS2
SARS is like AIDS, it's a phenotypic description of symptoms. SARS = Severe Accute Respriatory Syndrome. Covid was another example of "holy shit, this shit is crazy" ie 'Severe and Accute'. But covid wasn't the same as the first SARS. So it's just "SARS2". But they are different. The SARS virus creates the SARS symptoms. Covid also creates SARS symptoms, but it's not the SARS virus so you gotta call it something different. So we call it SARS2, but media got confused and heard it called covid-1 (Corona virus (CoV) ID 2019) and ran with it. But it's just another corona virus creating SARS symptoms. SARS2. Both corona viruses though, so similar enough with enough conserved mechanisms that what works for one might(?) work for the other.
But you don't know unless you try. Validating an existing approach that works for one coronavirus to demonstrate it works on the other virus is important.
An assay is just a fixed, routine experiment designed to generate similar repeatable results. Like running an alamar blue assay to return drug toxicity, or a custom assay designed to evaluate drug efficacy by surrogate (entry inhibition, binding inhibition, assembly inhibition, etc). What's important is validating that the results are represntative, that they are likely to reproduce when you go back to a larger model.
As for throughput, simply imagine doing an assay in cell culture with every control, drug, in triplicate, in their own petri dish. Imagine hundreds of dishes filling your hood. Imagine the amount of media that would take, the incubator space, the time to pipette individually moving from one step to the next... and then imagine instead doing so in a 384 well plate with a multi channel pipette. Imagine not needing to involve the sensitive conditions of cells because the assay is de novo. Speed goes up two orders of magnitude, cost goes down an order of magnitude.
But is that de novo assay representative of what you'd have gotten in cell culture? And is the cell culture result representative of what you'd see in animal? And is the animal model representative of what you'll get in humans? This issue is arguably THE issue in medical research, so assessing how representative something is a huge part of the process.
Demonstrating that your animal model reproduces the same pathology as in humans is critical, or what works in animals won't work in humans.
Demonstrating that your cell line can replicate the whole life cycle of the virus with no skipped or altered steps is critical if you're going to test in vitro.
Demonstrating that your high throughput assay replicates the specific isolated mechanism/interaction is critical for testing de novo/in silica.
There's lots of ways to do that. But what machines you use and what you measure depend a lot on what you're doing. Binding assays could be western blot and 384
So lets say you create a binding assay to evaluate how effective drugs are in inhibiting that binding (just an example). How do you test that the binding you're evaluating is the same as the binding the actual virus is involved it? Well, a great way would be to measure a number of variables of your surrogate and compare it to the genuine article. Do they match up, or is something off? And if something is off, what can you change until the surrogate matches the real thing?
But to do that, you need the real thing. Irradiated (inactivated, the gentic material has been destroyed, no risk of infection) to test against. This is a service the CDC provides. It's how labs like mine can work with things like Ebola without risk, without the cost and training that's required in running a BSL-3+ lab with clean rooms and bunny suits.
But the CDC wouldn't even retrun our inquiries. Which is odd. So I never got the samples I needed. So is the assay which is (probably) representative for SARS1 also representative for SARS2? Will the drugs I find in that assay likely to repeat in cell culture, animals, and humans? Well, I can't know. And that's a problem. So if the CDC gave a shit about the drug discovery process for covid, this would be among their priorities. That it ISN'T part of their priorities says everything. They are sufficiently co-opted to just be a vaccine 'health as a subscription service' shill organization.
That's good to know. Might I ask if you know what machines do you use for validating the assay and what corporation are they from? I have a strong suspicion against Illumina though I don't have much to go off on.
I agree, I do not like the mystical conspiracies you'll sometimes see here either.
I may be wrong on some things and I want to know them so I can learn and progress towards truth.
I am hesitant to detail the assay I would be using the particles as validation for as it would likely be enough to out me and my lab and living where I live has instilled in me an odd mix of caution for precisely when and where and how I stick my neck out.
In general terms, drug discovery requires a degree of throughput that can't be accomplished by doing things the 'best' way. One can't for example, infect tens of thousands of monkeys with SIV to test individual drugs against - ethically you've gone from a grey area to something utterly reprehensible. Doing so would also take a gargantuan amount of time and cost astronomical amounts.
It's better to do things in cell culture. Better throughput, even though as a collection of cells rather than a living breathing immune system or integrated organs it might not be quite as representative. Do the results in cell culture transfer to animal? Often not.
But even cell culture is low throughput, so in reality the only way to have a shot is to design assays which approximate the functions of relevance in the cells that are representative of the larger organism to test agaisnt the mechanism of action you're trying to develop against - for example, merely replicating the receptor binding that is prelude to entry, or the binding regions of a capsif they rely on for assembly.
But how do you know your assay which so removed from the integrated whole is actually representative? Well, you validate it. And for many such examples, trialing against actual legitimate viral particles is enough. If the virion binds the same as your stand in, and shows similar reactivity to some of your early drug candidates, you can have enough confidence (never needs to be 100%) that's it's then worth the time to roll the dice running that assy hundreds of times on tens of thousands of unique compounds to find one or two worth doing more intensie looks on...
...In cell culture. And if that works, in Animal trials. So as you could imagine, I am happy to receive other irradiation particles from the CDC, and a little perterbed when I don't know if my generic SARS1 coronavirus assay is sufficiently representative to SARS2 to be worth the time.
I swear that wasn't supposed to be that long.
Dude I'm totally good with it, I'm at a university and I have some (albeit very very small) experience with what you're doing.
I'm not as interested in the assay but the equipment (or really the manufacturer) that measures everything.
(Correct me if I'm wrong) an assay is a substance that you get that is analyzed for having our Covid-19 virus.
Throughput is the rate of production or the rate at which something is processed, so a cell culture gives little data to process but if you design the cell culture (collection of cells) around the cells the Covid-19 virus is supposed to drastically infect and alter, and if the assay (which is supposed to have the virus) binds to the cells the same way Covid-19 virus does.
If the virus (idk difference between virus and viron) binds the same way, you revalidate it with other cells and once you've sufficiently done that, you have the confidence that this assay is worth the time to run hundreds of times on tens of thousands of samples/compounds to see maybe one or 2 compounds that react differently (maybe for vaccine or interesting study).
What are irradiation particles?
Also it looks like you cut off towards the end, please tell me more when you get the chance! So your generic SARS1 coronavirus didn't represent SARS2? That's really weird, they should be practically the same thing with a better binding mechanism for SARS2
SARS is like AIDS, it's a phenotypic description of symptoms. SARS = Severe Accute Respriatory Syndrome. Covid was another example of "holy shit, this shit is crazy" ie 'Severe and Accute'. But covid wasn't the same as the first SARS. So it's just "SARS2". But they are different. The SARS virus creates the SARS symptoms. Covid also creates SARS symptoms, but it's not the SARS virus so you gotta call it something different. So we call it SARS2, but media got confused and heard it called covid-1 (Corona virus (CoV) ID 2019) and ran with it. But it's just another corona virus creating SARS symptoms. SARS2. Both corona viruses though, so similar enough with enough conserved mechanisms that what works for one might(?) work for the other.
But you don't know unless you try. Validating an existing approach that works for one coronavirus to demonstrate it works on the other virus is important.
An assay is just a fixed, routine experiment designed to generate similar repeatable results. Like running an alamar blue assay to return drug toxicity, or a custom assay designed to evaluate drug efficacy by surrogate (entry inhibition, binding inhibition, assembly inhibition, etc). What's important is validating that the results are represntative, that they are likely to reproduce when you go back to a larger model.
As for throughput, simply imagine doing an assay in cell culture with every control, drug, in triplicate, in their own petri dish. Imagine hundreds of dishes filling your hood. Imagine the amount of media that would take, the incubator space, the time to pipette individually moving from one step to the next... and then imagine instead doing so in a 384 well plate with a multi channel pipette. Imagine not needing to involve the sensitive conditions of cells because the assay is de novo. Speed goes up two orders of magnitude, cost goes down an order of magnitude.
But is that de novo assay representative of what you'd have gotten in cell culture? And is the cell culture result representative of what you'd see in animal? And is the animal model representative of what you'll get in humans? This issue is arguably THE issue in medical research, so assessing how representative something is a huge part of the process.
Demonstrating that your animal model reproduces the same pathology as in humans is critical, or what works in animals won't work in humans.
Demonstrating that your cell line can replicate the whole life cycle of the virus with no skipped or altered steps is critical if you're going to test in vitro.
Demonstrating that your high throughput assay replicates the specific isolated mechanism/interaction is critical for testing de novo/in silica.
There's lots of ways to do that. But what machines you use and what you measure depend a lot on what you're doing. Binding assays could be western blot and 384
So lets say you create a binding assay to evaluate how effective drugs are in inhibiting that binding (just an example). How do you test that the binding you're evaluating is the same as the binding the actual virus is involved it? Well, a great way would be to measure a number of variables of your surrogate and compare it to the genuine article. Do they match up, or is something off? And if something is off, what can you change until the surrogate matches the real thing?
But to do that, you need the real thing. Irradiated (inactivated, the gentic material has been destroyed, no risk of infection) to test against. This is a service the CDC provides. It's how labs like mine can work with things like Ebola without risk, without the cost and training that's required in running a BSL-3+ lab with clean rooms and bunny suits.
But the CDC wouldn't even retrun our inquiries. Which is odd. So I never got the samples I needed. So is the assay which is (probably) representative for SARS1 also representative for SARS2? Will the drugs I find in that assay likely to repeat in cell culture, animals, and humans? Well, I can't know. And that's a problem. So if the CDC gave a shit about the drug discovery process for covid, this would be among their priorities. That it ISN'T part of their priorities says everything. They are sufficiently co-opted to just be a vaccine 'health as a subscription service' shill organization.