Dumb question conjugating proteins

[Kylonicus]

I know this is a dumb question, but how do you conjugate/combine proteins?

 

 

The reason I wanna know, is that I need a large quanity of a very expensive antibody for a piece of research I am doing. If I can find out how to conjugate proteins then, I can simply acquire the original substance, conjugate it to a viral protein, and then make a sub-type vaccination, and produce antibodies from that.

 

Could anybody tell me?

[Kylonicus]

Is anybody going to reply?

[donkey]

doesn't look like it

[Bluenoise]

Never done this but I'd assume you'd combine the two coding regions from the mRNA, clone it express it and purify?

Doing it chemically would probably very tricky.

 

Have you tried just producing the antibody from the non-conjugated protein? You should still be able to get an immune response if it's placed in a forien host.

[Kylonicus]

If the protein source was mouse, and I wanted to inject it into another mouse, do you think it would probably still develop an antibody to it?

[Bluenoise]

No it wouldn't. You would need to inject it into something else. Like a rabbit for instance.

[Bluenoise]

Yeah try conjugating it by combining the coding regions of the mRNA. It may work, it may not. I'd do some research before investing the time into it though.

Sorry for the double post.

[Kylonicus]

BTW, thanks for answering my questions Blue Noise.

 

If I were to inject NogoA from a mouse, into a rabbit, and there were differences, so that the rabbit developed an immunity to NogoA mouse version, would that cause the NogoA in the rabbit to be targeted? I am assuming it would, because part of the antibody would remain similar enough to the NogoA rabbit version, that it would attach itself to the similar portion causing it to have the same effects as if it had been originally innoculated.

 

Am I right?

[Bluenoise]

No I don't think it would.

 

I think the epitopes that the system would develope antibodies towards would probably be those that differ between the two species, so it should produce a unique enough antibody.

 

One problem that I can see with this approach is that both these species are rodents and maybe so simulairly related that you might get no immune reaction at all. Using a horse would be far more effective I think, but that's probably not very plausible for research.

 

Remember they used to use pig insulin in humans effectively.

[Kylonicus]

Then perhaps, if it were a horse, or some other large mammal, do you think that the antibody would be against the NogoA protein, that came originally from the rat?

[Bluenoise]

Yes if you put a mouse protein into a horse it would almost certainly produce antibodies against that mouse protein.

[Kylonicus]

BTW, THANK YOU, your answers are helping me ALOT,

 

The stuff I need to buy in order to do this is very expensive. Cost like 325 bucks per 50 ug.

 

But you think that the antibody might not target the horse protein, if they are very, very similar in structure?

[Bluenoise]

Yes , but it's very unlikely that they'll be that simulair. It's very typical to use rabbits and horses for antibody production to each other or humans since they're all quite different.

If you know the sequences of both the mouse and horse versions of this protein you can try to do a statistical analysis with bioinformatics tools. I doubt there would be any need to do this however, but it would only take a little while so I can't see any reason not to.

got to get off the computer I'll catch you later.

[Kylonicus]

Thanks for all your assistance Blue Noise, you've been a great help to me.

[scientistsahai]

know the sequences of both the mouse and horse versions of this protein you can try to do a statistical analysis with bioinformatics tools

 

 

Hi Quite interesting though ,but can anyone tell me which Bioinformatics tools would be helpful. Please list all and those that are available on the Internet

[Bluenoise]

http://www.ncbi.nlm.nih.gov/

The place to start.

Blast is very useful