Immunoaffinity Subtraction

Christer Ericsson christer.ericsson at mog.ki.se
Mon May 8 14:10:36 EST 1995


In article <3ohqli$795 at worak.kaist.ac.kr>, dykim at bioneer.kaist.ac.kr
(Dooyeon Kim) wrote:

> I have some ideas on the detection and purification of newly expressed
protein.
> Instead of using cDNA subtraction or 2D PAGE analysis, I will use immunosubt
> ractive methods. The princeple of this method is producing the polyclonal
> antibodies against whole protein mixture of tissue or cultured cell and select
> ing the antibodies against newly expressed protein by passing the antiserum
> against the protein(antigen) column of control tissue of cell line.
After immuno
> subtraction, the newly expressed protein will be easily purified by immuno
> precipitation and detected on SDS PAGE gels. Dose anybody advice me about this
> idea? Any Comments will be greatly appreciated. Thanks for reading

I have been toying with similar ideas for some time, but never got around
to trying it. I suspect one may have problems with some (many?) proteins
not being very immunogenic in a complex mixture (due to immunodominance or
whatever) and so after spending time and money on raising and depleting
antiserum, you may end up not being able to purify your protein. On the
other hand, it may be a new method that will revolutionize the way we
work?

Is there anybody out there who actually tried, and is willing to share
his/her experience?

Christer

-- 
Christer Ericsson, Dept. of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institute, S-171 77 Stockholm, Sweden.

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