Antibodies to recombinant proteins - Any advice on production of proteins? (Jun/13/2003 )
I am looking to have some polyclonal antibodies produced! in order for me to do this I need to have my proteins expressed. I have found a company based in the UK called Harlan Sera-Lab, who can produce my recombinant proteins from the gene and have my polyclonal with me within 15wks. Would you say this is my best route? the company sound very reasonable on price and have told me that if they did fail to express any of my proteins I would not be charged for their efforts. I have never expressed a protein myself but my colleague tells me that this service sounds very good, if anyone can let me know their thoughts it would be great.
Kind regards
Peter Davis
Hi there!
In brief: If you can afford it, do it!
I myself had to express a protein, which can be relatively easy, if you're lucky. Most of the time you're not, as in my case .... (wrong codon usage for your exprssion systen, little or no expression, toxic products etc.)
Then you have to do a lot of try and error in expression (different systems, different concentrations of inductors, different times of induction etc.). So when you get your protein expressed at reasonable levels, it's time for purification. There can go anything wrong with that, too.
And then, finally you're in for the immunisation. So you have to get youself an anmial, preferably a big one (bigger animal = more blood, but bigger animals = bigger teeth). Usually animal expriments have to be aprooved of by some official commitee or something, wich is more or less difficult.
Then, it's immunisation time. Different routes for administration for the antigen may have to be checked, different adjuvates, maybe different animals.... And animals sometimes are completely uncooperative when you try to hurt them with a pointy sharp objects.
After some time, it's bloodletting time. Again, animals can be very uncooperative... (see above)
When done, you need to test your sera, and if you're lucky, there's some activity....
So, in summary, it's a lot of work to generate a polyclonal serum, and if you're lucky, it works. But if Harlan offers you some kind of money back-guarantee that their antibodies will do the job, and you have the money - DO IT!
Mike
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Never give up, never surrender!
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Hi Dynamite,
This is only my personal opinion but I would take up Harlan's offer, I actually have a colleague that used them a little while ago and they were excellent.
We actually had a real problem expressing our protein and went to Harlan because they offered the no charge on the protein if they failed to express it.
They had no such problem and we ended up with a very nice polyclonal antibody for a pretty affordable price.
I know we would use them again, there are probably some other companies out there that could do the job but you know what it's like you tend to stick with what you know.
Thought Mikes answer was brilliant!!! made me chuckle.
Take care and good luck with your protein and whatever you decide to do.
Regards
Chris