The generic vs pharma GH discussion

[BamaCrazy]

Status?

I was planning on getting some more Serostim so I could run head to head against a "premium" UG GH and a typical UG "generic" GH. The "premium" supposedly tests better against pharma standard with HPLC-MS. I don't have that data and vendor won't share it.

The price Jano quoted me was 400 USD for a pair wise comparison with CD and 600 USD for 3 samples. He stated this was no markup on his part with outside lab. I want to speak with this lab first if we go with CD to confirm near vs far UV and make sure it isn't a mess like the oil based endotoxin analyses.

If someone orders NMR then you are on your own as Jano doesn't have the bandwidth to interpret results and that ain't my thing. Do we have NMR guru here?

Hence CD is probably best incremental test we could pursue IMO. Interpretation is simple if you get the testing conditions right.

Fyi.

Thoughts/feedback?

I’m in once you all are ready.

 

[Spaceman Spiff]

ill throw some money in

 

[readalot]

Tools and methods for circular dichroism spectroscopy of proteins: a tutorial review

Circular dichroism (CD) spectroscopy is a widely-used method in biochemistry, structural biology and pharmaceutical chemistry. More than 24 000 papers published in the past decade have included CD characterisations of proteins; many of those studies have also included other complementary...

pubs.rsc.org

Full paper.

 

[Ghoul]

Tools and methods for circular dichroism spectroscopy of proteins: a tutorial review

Circular dichroism (CD) spectroscopy is a widely-used method in biochemistry, structural biology and pharmaceutical chemistry. More than 24 000 papers published in the past decade have included CD characterisations of proteins; many of those studies have also included other complementary...

pubs.rsc.org

Full paper.

An important step forward.

It's important to wait after reconstitution for the greatest strengths of CD to be utilized.

Perhaps come up with a formula for average user "shelf time".

Ideally something like 1 day per 3iu before analysis. IE 4 days for a 12iu via, 12 for a 36iu vial.

Or if that's too complicated logistically, reconstitute and wait a week.

Also, a fixed volume for reconstitution. 2ml might be a good middle ground.

 

[Spaceman Spiff]

L

An important step forward.

It's important to wait after reconstitution for the greatest strengths of CD to be utilized.

Perhaps come up with a formula for average user "shelf time".

Something like 1 day per 3iu before analysis.

no discussion of submission for actual testing. Got it.

No money to put in, zilch, nada.

 

[readalot]

Going to get the pharma standard (Sero) unless someone else has some. I got all the other samples. Easier for one person to submit everything. @Sampei said he would organize so as trusted member perhaps he can collect the money?

Unless others dont want Sero as the standard. If so, Jano said he could provide reference standard for 300 usd. Obviously 1 vial of Sero is cheaper than that... $625/7.

Just let me know everyone's preference.

 

[BamaCrazy]

Going to get the pharma standard (Sero) unless someone else has some. I got all the other samples. Easier for one person to submit everything. @Sampei said he would organize so as trusted member perhaps he can collect the money?

Unless others dont want Sero as the standard. If so, Jano said he could provide reference standard for 300 usd. Obviously 1 vial of Sero is cheaper than that... $625/7.

Just let me know everyone's preference.

I’m ready to support whatever the best comparison is. I’m certain we can find the funding.

 

[Diixencider]

Going to get the pharma standard (Sero) unless someone else has some. I got all the other samples. Easier for one person to submit everything. @Sampei said he would organize so as trusted member perhaps he can collect the money?

Unless others dont want Sero as the standard. If so, Jano said he could provide reference standard for 300 usd. Obviously 1 vial of Sero is cheaper than that... $625/7.

Just let me know everyone's preference.

i got some coin for science lmk

 

[readalot]

I’m ready to support whatever the best comparison is. I’m certain we can find the funding.

$$$ aint the issue. I agree. I'm psyched members want to do a group project. I was planning to do it myself but great we can bring folks together. I know I need to do a much better job on that front. I have listened to the feedback.

And if folks disagree on the CD and want NMR I'll respect that. Just providing my (0.02) cents.

Thanks.

 

[Diixencider]

$$$ aint the issue. I agree. I'm psyched members want to do a group project. I was planning to do it myself but great we can bring folks together. I know I need to do a much better job on that front. I have listened to the feedback.

And if folks disagree on the CD and want NMR I'll respect that. Just providing my (0.02) cents.

Thanks.

I just wana chip in so we can get the answers we all want. generic vs pharma is always the argument

 

[readalot]

An important step forward.

It's important to wait after reconstitution for the greatest strengths of CD to be utilized.

Perhaps come up with a formula for average user "shelf time".

Ideally something like 1 day per 3iu before analysis. IE 4 days for a 12iu via, 12 for a 36iu vial.

Or if that's too complicated logistically, reconstitute and wait a week.

Also, a fixed volume for reconstitution. 2ml might be a good middle ground.

Appreciate the feedback. Gotta get with the lab and make sure I dont screw it up this time. Very Nice of @janoshik to find a lab for us and it ain't like this gets done every day in the UG. Usually takes a few attempts to smooth out the kinks.

 

[readalot]

I just wana chip in so we can get the answers we all want. generic vs pharma is always the argument

Its definitely a step in the right direction. Combined with the Nb2 bioassay it would be a useful evolution in GH testing IMO.

 

[readalot]

GH testing

UG GH testing.

 

[AlexDavis43]

I just wana chip in so we can get the answers we all want. generic vs pharma is always the argument

I see no one replied yet. Probably because this testing will not tell if UGL or pharma is better.

If UGL GH was misfolded or whatever, people wouldn't be getting any of the expected results from it.

 

[Diixencider]

I see no one replied yet. Probably because this testing will not tell if UGL or pharma is better.

If UGL GH was misfolded or whatever, people wouldn't be getting any of the expected results from it.

yeah ive seen stellar bloodwork with ugl gh so the markup for pharma shit is to rich for my blood lol

 

[Spaceman Spiff]

I see no one replied yet. Probably because this testing will not tell if UGL or pharma is better.

If UGL GH was misfolded or whatever, people wouldn't be getting any of the expected results from it.

Agreed... Been talking about NMR this whole time and won't do jack shit to get the test done

 

[Ghoul]

Appreciate the feedback. Gotta get with the lab and make sure I dont screw it up this time. Very Nice of @janoshik to find a lab for us and it ain't like this gets done every day in the UG. Usually takes a few attempts to smooth out the kinks.

This is your show man, but you may find this useful. In discussion with the guys that handled the Tirz NMR analysis, they offered CD as an option for rHGH that would capture defects they thought likely in UGL rHGH, but be cost effective enough for routine analysis. (unlike NMR)

Both of these conditions still appear "pure" under HPLC-MS.

1. rhGH is relatively “sticky” once partially unfolded. Many forms of stress induce this partial unfolding (IE less than ideal storage conditions like uncontrolled temps in transport, and the haphazard formulations common in UGL). It can still be bioactive in this condition. Once partially unfolded it's susceptible to self‐associating into inactive aggregates. This can happen seconds after reconstitution (likely the cause of "cloudiness") or over days or weeks.

For partial unfolding they'll need to look for "A reduction in the characteristic 208/222 nm negative minima (loss of α‐helix signal) and/or the appearance of a β‐sheet–like shoulder around ~216 nm."

So this is a measure of quality that predicts how quickly the rHGH will degrade. Monitoring by far-UV CD (for α-helix loss) is a sensitive way to catch it early.

--------

2. Although well manufactured rhGH is stable, there's plenty of room for sloppiness in UGL rHGH manufacturing via slight deviations such as transient pH shifts during dialysis, small amounts of organic solvent carryover, or buffer ionic‐strength change which can damage the hydrogen‐bond network in its core helices. Instead of completely aggregating, many molecules simply lose one or two helices and adopt a molten-globule–like state.

For this, they'll need to look for "A shallower double minimum at 208 nm and 222 nm compared to a “textbook” rhGH spectrum. If only a fraction of molecules misfold, you see a blend (native + unfolded) and can estimate the percentage of misfolded species."

This will find inactive rHGH that hasn't aggregated but's inactive despite appearing pure on HPLC-MS. The next best thing to an bioactivity assay.

We couldn't do this because it's a US based lab and in the end there was no legal workaround that would allow them to accept a rHGH sample from us.

Will be great to follow your progress regardless of the process you settle on.

Good luck!

 

[BamaCrazy]

This is your show man, but you may find this useful. In discussion with the guys that handled the Tirz NMR analysis, they offered CD as an option for rHGH that would capture defects they thought likely in UGL rHGH, but be cost effective enough for routine analysis. (unlike NMR)

Both of these conditions still appear "pure" under HPLC-MS.

1. rhGH is relatively “sticky” once partially unfolded. Many forms of stress induce this partial unfolding (IE less than ideal storage conditions like uncontrolled temps in transport, and the haphazard formulations common in UGL). It can still be bioactive in this condition. Once partially unfolded it's susceptible to self‐associating into inactive aggregates. This can happen seconds after reconstitution (likely the cause of "cloudiness") or over days or weeks.

For partial unfolding they'll need to look for "A reduction in the characteristic 208/222 nm negative minima (loss of α‐helix signal) and/or the appearance of a β‐sheet–like shoulder around ~216 nm."

So this is a measure of quality that predicts how quickly the rHGH will degrade. Monitoring by far-UV CD (for α-helix loss) is a sensitive way to catch it early.

--------

2. Although well manufactured rhGH is stable, there's plenty of room for sloppiness in UGL rHGH manufacturing via slight deviations such as transient pH shifts during dialysis, small amounts of organic solvent carryover, or buffer ionic‐strength change which can damage the hydrogen‐bond network in its core helices. Instead of completely aggregating, many molecules simply lose one or two helices and adopt a molten-globule–like state.

For this, they'll need to look for "A shallower double minimum at 208 nm and 222 nm compared to a “textbook” rhGH spectrum. If only a fraction of molecules misfold, you see a blend (native + unfolded) and can estimate the percentage of misfolded species."

This will find inactive rHGH that hasn't aggregated but's inactive despite appearing pure on HPLC-MS. The next best thing to an bioactivity assay.

We couldn't do this because it's a US based lab and in the end there was no legal workaround that would allow them to accept a rHGH sample from us.

Will be great to follow your progress regardless of the process you settle on.

Good luck!

So once again all talk, no action. Correction, all ChatGPT cut and paste, no action.

 

[Spaceman Spiff]

So once again all talk, no action. Correction, all ChatGPT cut and paste, no action.

Folding, stickied, who gives a shit. While igf1 doesn't mean everything, seems like people are still getting a solid response.

This fucking clown keeps finding a way to downplay testing but won't do any of his own.

Then he cries like a little bitch saying we are picking on him.

I am glad he is proving our point. Theoretical bullshit with no data collection

 

[bananafeet]

Why do the arguments for pharma HGH keep getting more and more strange.

First it was purity, then it was other additives and now it's protien shape? Or misfolded proteins.

Seems like people are clutching at straws.

Or am I missing something? If something tests as sugar it's sugar right?