Fluorescent protein uptake?

[schudini]

A rather technical but probably interesting question.

Would the consumption of organisms containing a fluorescent protein eventually lead to uptake and incorporation of the protein, or just result in fluorescent detritus?

 

[TheH]

It's unlikely that the protein would remain undigested and retain it's fluorescence except perhaps for a few moments. Although it has been hypothesized that some species steal fluorescent proteins (in the deep ocean), they would be highly specialized.

 

[schudini]

That's what I was thinking too.

 

[WLachnit]

A fluorescent protein will be degraded by enzymes just like any other protein and therefore lose it's flourescent properties before it even becomes detritus. However, organisms can be made to fluoresce using fluorescent proteins, but this requires genetically engineering them to do so. In fact, by using green fluorescent protein from jelly fish, this is how fluorescent mice (and other organisms) are made.

 

[schudini]

<a href=showthread.php?s=&postid=14993613#post14993613 target=_blank>Originally posted</a> by WLachnit
A fluorescent protein will be degraded by enzymes just like any other protein and therefore lose it's flourescent properties before it even becomes detritus. However, organisms can be made to fluoresce using fluorescent proteins, but this requires genetically engineering them to do so. In fact, by using green fluorescent protein from jelly fish, this is how fluorescent mice (and other organisms) are made.

That's one way to do it. I was thinking of a more non-invasive technique, possibly using fluorophores or quantum dots.

What type of "biotech" is your specialty?

 

[WLachnit]

<a href=showthread.php?s=&postid=14993922#post14993922 target=_blank>Originally posted</a> by schudini
That's one way to do it. I was thinking of a more non-invasive technique, possibly using fluorophores or quantum dots.

Well, you would have to use a fluorophore that is taken up by the cells of the organism. Quantum dots, while small, would have to be phagocytosed I would think to be taken up cells. The other way to do it is to infect the organism by using a gentically engineered virus that encodes the fluorescent protein suitable for the host. The host will then incorporate it epigenetically and express it for a while.

<a href=showthread.php?s=&postid=14993922#post14993922 target=_blank>Originally posted</a> by schudini
What type of "biotech" is your specialty?

Over the years, everything from drug discovery to the development of tools for life science research including ones just described.

 

[schudini]

<a href=showthread.php?s=&postid=14994662#post14994662 target=_blank>Originally posted</a> by WLachnit
Well, you would have to use a fluorophore that is taken up by the cells of the organism. Quantum dots, while small, would have to be phagocytosed I would think to be taken up cells. The other way to do it is to infect the organism by using a gentically engineered virus that encodes the fluorescent protein suitable for the host. The host will then incorporate it epigenetically and express it for a while.

I think I will stay away from genetically engineered viruses. Too many unknowns.

Over the years, everything from drug discovery to the development of tools for life science research including ones just described.

Interesting. Do you have experience using MALDI-TOF MS for protein/peptide detection? I'm working on a technique for sensitivity enhancement.

 

[lbaldrey]

WTbeep!

That just went over my head

 

[WLachnit]

<a href=showthread.php?s=&postid=14995634#post14995634 target=_blank>Originally posted</a> by schudini
Interesting. Do you have experience using MALDI-TOF MS for protein/peptide detection? I'm working on a technique for sensitivity enhancement. [/B]

I am fairly familar with the technique but, have no personal experience with it. Actually, currently commercializing a platform for protein isoform detection with attomole sensitivity.

I would be very curious about your sensitivity enhancement technique.

 

[sowellj]

totally off topic ... but I do a good bit of triple quad work and some proteomics (TOFs and traps). If you know the peptide you are after, a triple quad is likely your best bet, i.e. targeted proteomics.

 

[schudini]

<a href=showthread.php?s=&postid=14998275#post14998275 target=_blank>Originally posted</a> by WLachnit
I am fairly familiar with the technique but, have no personal experience with it. Actually, currently commercializing a platform for protein isoform detection with attomole sensitivity.

I would be very curious about your sensitivity enhancement technique.

Attomole sensitivity is good, but how robust is the preparation (i.e. required precision for reproducibility at that concentration)? What is the required experience level for the operator?

My technique utilizes carbon nanotubes (my background) to influence matrix solution behavior. Only peptides have been tested thus far, however we feel that applications in protein and DNA detections could benefit from the technology. Since we have applied for a patent, I will have to check before I can disclose additional information.

 

[schudini]

<a href=showthread.php?s=&postid=14998500#post14998500 target=_blank>Originally posted</a> by sowellj
totally off topic ... but I do a good bit of triple quad work and some proteomics (TOFs and traps). If you know the peptide you are after, a triple quad is likely your best bet, i.e. targeted proteomics.

If you are referring to the original post about fluorescent proteins or fluorophores, I don't have a particular organism in mind to study. Rather just a passing idea.

Also, I don't think that I have access to any triple quads, however we are building a medium vacuum single quad with electrospray injection for liquids (different project than previous post).