15 replies to this topic

[Plasticperson]

Hello Longecity, 

 

I have decided to pursue the sequencing of peptides in cerebrolysin. To accomplish the sequencing I'm hiring a highly qualified lab that regularly analyzes protein sequences. The lab I intend on using is called Alphalyse. Here is a link (http://www.alphalyse...tification.html) that dictates their protein identification information. Im going to be using a modified process of the MALDI-MS/MS or nanoLC-MS/MS (Last Paragraph). The price of sequencing is $350 (MALDI) and $600 (nanoLC). I'm in contact with the company, they're fully aware that I'm not yet sure of the amount of peptides/proteins within cerebrolysin. Subsequently, the amount of different proteins in a sample determines which process is used. I'm most likely going to be forced into choosing the nanoLC even though it might not be necessary. Once I send out the sample the turn around time for the sequences would be 2-8 days. Additionally, not only will the lab provide the sequences but also the presumed proteins that the sequences belong to.

 

I'm going to share the sequences and presumed proteins with longecity   . However, I have a favor to ask the Longecity community. Since I am spending a substantial amount of money on the sequencing and I don't have any cerebrolysin at my immediate disposal I would appreciate someone donating a single vial of cerebrolysin. Any sized vial would work. I'm sorry if this comes off as a selfish request that is not my intention. Buying a pack of vials would put me over my budget and waiting till it arrives from Europe would lengthen the whole process(I live in US). I can ensure you this isn't a scam to receive a free vial of cerebrolysin. PM me and we can further discuss details. 

 

Lastly, I'd like to go over the modified process of the sequencing. Usually whole proteins are submitted for analysis and broken down into peptides with an enzyme called lysate. However, cerebrolysin has already been broken down into peptides. Most likely by the enzyme lysate hence the term cerebroLYSIN. Since cerebrolysin is already broken down, a part of the process is already taken care of. 

 

Please take my dontation  request into consideration it would help me out a lot. Also knowing the peptides would be soo beneficial and just all around interesting.

 

 

Thanks,

Plasticperson

[Flex]

Cudos, a very good Idea.

But I wouldn´t rely too much on the hope for the donations...sigh

Maybe it would be better to gather first the donations.

 

I dont know if You heard that, but one user said that he read the Patent of Cerebrolysin and found 3 peptides to be responsible for the effects.

This would make sence, because the inventor of Cerebrolysin "should" have done resarch on their own, to find out the responsible substances.

If this is true, then You dont have to make the test.

 

However, when I looked trough the Patent I couldn´t find anything regarding the 3 peptides

http://www.google.co...0452299A1?cl=en

 

Nevertheless, I´m hoping to get to know the responsible peptides.

 

 

 

 

[malbecman]

 Hi Plasticperson,

 

 I run an academic mass spectrometry facility that does this kind of analysis for our campus.   That price is a little high, IMHO and there are other core facilities that could

probably do this for you.   How many peptides are present in cerebrolysin?  I did a little Pubmed searching and came up with this "Cerebrolysin is a porcine brain-derived preparation of low-molecular-weight neuropeptides (10 kDa) and free amino acids that shows pharmacodynamic properties similar to those of naturally occurring neurotrophic factors."

 

 If it's a really complex mixture, it might be difficult for Alphalyse or anyone to get complete sequences for you.  The other components in the mixture can suppress the signals of the peptides.   Are they charging you per sequence or does that price include ALL the sequences that they can find for you?

[Plasticperson]

Cudos, a very good Idea.

But I wouldn´t rely too much on the hope for the donations...sigh

Maybe it would be better to gather first the donations.

 

I dont know if You heard that, but one user said that he read the Patent of Cerebrolysin and found 3 peptides to be responsible for the effects.

This would make sence, because the inventor of Cerebrolysin "should" have done resarch on their own, to find out the responsible substances.

If this is true, then You dont have to make the test.

 

However, when I looked trough the Patent I couldn´t find anything regarding the 3 peptides

http://www.google.co...0452299A1?cl=en

 

Nevertheless, I´m hoping to get to know the responsible peptides.

 

 

 

 

The chance of getting a vial donation would probably be very low :( but i figured it would be worth a try. I'm sure we will know the peptides in cerebrolysin in due time i'm really interested in getting the testing and have the extra money. 

 

 Hi Plasticperson,

 

 I run an academic mass spectrometry facility that does this kind of analysis for our campus.   That price is a little high, IMHO and there are other core facilities that could

probably do this for you.   How many peptides are present in cerebrolysin?  I did a little Pubmed searching and came up with this "Cerebrolysin is a porcine brain-derived preparation of low-molecular-weight neuropeptides (10 kDa) and free amino acids that shows pharmacodynamic properties similar to those of naturally occurring neurotrophic factors."

 

 If it's a really complex mixture, it might be difficult for Alphalyse or anyone to get complete sequences for you.  The other components in the mixture can suppress the signals of the peptides.   Are they charging you per sequence or does that price include ALL the sequences that they can find for you?

 

The price for the nanoLC includes between 5-50 proteins which are then fractioned into numerous peptides. However, cerebrolysin has already been fractioned. I don't know the exact amount of peptides in the mixture which is a problem. Ive read some people guessing between 10-19 all the way up to hundreds.. its really hard to tell. Also, would the amino acid mixture affect the sequencing? That was one of my worries but i figured that the liquid chromatography would be able to separate the peptides from the amino acids. 

 

Alphalyse didn't really seem to think of the amount peptides as a problem however I don't know if I'm talking to one of their trained chemists or sales representative. I'm going to call them today to discuss the topic. 

 

Also do u know any other testing facilities off the top of your head? I picked Alphalyse because they seemed very experienced and I didn't know the exact amount of peptides in cerebrolysin. However, if i could find a facility that is just as competent and has a lower price I would definitely contact them to discuss cerebrolysin.

Edited by Plasticperson, 06 May 2014 - 07:21 PM.

[Plasticperson]

Great news

 

Just talked to an Alphalyse chemist on the phone they saw NO PROBLEMS with the cerebrolysin mixture as long as a the nanoLC option was picked. Amino acids wouldn't pose a problem and neither would the amount of peptides. Apparently some single proteins can break up into hundreds of peptides. 

 

Now if someone could donate a single vial of cerebrolysin we could get on with the analysis.. c'mon longecity I have faith in you   

Edited by Plasticperson, 06 May 2014 - 07:55 PM.

[formergenius]

I have a 10ml ampoule you can have, but I'm in Europe..

[malbecman]

Glad to hear they did not think it would be a technical problem to do that number of peptides for you. Hopefully you'll get a vial donated soon.

[Mr Matsubayashi]

I have some expired product, I'm sure it will suit your purposes fine. PM me if you're interested.

 

Scrap that, the countries post system will not allow it.

 

Edited by Mr Matsubayashi, 19 May 2014 - 11:22 AM.

[OpaqueMind]

I've already sent one to him and the mail trace site says it's been received, so hopefully we'll have the sequence soon! Just letting you guys know so you don't have to offer yours up any more.

[FW900]

I've already sent one to him and the mail trace site says it's been received, so hopefully we'll have the sequence soon! Just letting you guys know so you don't have to offer yours up any more.

 

Thanks for posting this and thanks being the one to donate the ampule to Plasticperson! I was about to offer a 1 ml Cerebrolysin ampule. In the event that yours did arrive damaged (unlikely, the ampules seem to take a beating) I'd still be happy to send the 1ml ampule to Plasticperson.

 

Plasticperson, this is truly a worthwhile contribution, thank you for taking the time and expense to have the peptides' sequences in Cerebrolysin determined!

[Plasticperson]

The company sequencing the peptides needs to know what cerebrolysin is dissolved in and if any/what buffers are present. Sequencing of 10kDa peptides is a bit more complicated that the company made it out to be. Is there anyway i can get this information? I really want this to happen.

[Keizo]

NaOH and water, I believe. Anything else I don't know, except that free amino acids are present besides the peptides ( http://wiseyoung.wor...2009/02/10/271/ )  You could always email:  office at everpharma.com if nothing else

Edited by Keizo, 11 June 2014 - 07:03 AM.

[Keizo]

http://www.longecity...-10#entry424977

[Plasticperson]

they want the enzymes used to fraction the proteins found in cerebrolysin 

[Wave Txlitl Met]

Each ml of this drug contains 3.00 mg of alanine, 0.25 mg of arginine, 3.00 mg of asparaginic acid, 0.06 g of cystine, 4.30 mg of glutaminic acid, 1.50 mg of glycine, 1.30 mg of histidine, 2.00 mg of isoleucine, 6.00 mg of leucine, 0.50 mg of methionine, 2.00 mg of phenylalanine, 2.00 mg of proline, 0.30 g of serine, 0.30 g of threonine, 0.50 g of tryptophane and 2.00 mg of thyrosine as amino acids as well as peptide having a molecular weight of 10.000 or less. In patent cited above.

[Wave Txlitl Met]

The amounts of each amino acid show three figures in mg for Serine, Threonine,  and Tryptophan. The other figures fail to exceed 4.3mg except Cysteine, which is 60mg. 

 

It may not be in the patent, but any person making a knock-off would use those three I've mentioned, and maybe Cysteine. Cysteine is 20% of serine or threonine and less than 15% of tryptophan. Would you need it? (rhetorical)