Re: Purification of the antipeptide antibody

Roger Murphy (murphy_r@licre.ludwig.edu.au)
Mon, 07 Sep 1998 09:13:40 +1000

At 01:37 PM 04-09-98 -0500, you wrote:
>Hi everyone,
>
>I need some advice. How many ways can one purify the antibody. Is coupling
>the synthetic peptide (not the peptide-KLH) to CNBr-activated Sepharose-4B
>and affinity purify the antibody the best way ? How about purifying the
>antibody by ammonium sulfate, DEAE-Affi-gel blue or protein A. Is the IgG
>fraction better antibody. Thanks.
>
>Bob Lee, UIC
>

Bob,

You need to be more specific about your antibody - is it polyclonal or
monoclonal? What do you want to use it for? Radioimmunoassay and
immunohistochemistry are usually okay (or even better) with polyclonal
serum, diluted appropriately and controlled for by absorbing out
non-specific interference from other immunoglobulins. Monoclonals are
better being purified, and the best way we've found is protein A or protein
G, followed by ion exchange and gel filtration....it's difficult to be more
specific without more details...

Cheers,

Roger

Roger Murphy, Ph.D.
Biological Production Facility
Ludwig Institute for Cancer Research
Austin & Repatriation Medical Centre
Studley Road,
Heidelberg, Vic. 3084
Australia.

Tel 61-3-94965463
Fax 61-3-94965436
Email murphy_r@licre.ludwig.edu.au