Affinity Chromatography

[Salih KAFKAS]

Hi,
Binding of lectins onto carbohydrate-based chromatography sorbents
(Sephadex, Sepharose etc.) seems to be unpredictable. So I would
recommend to use only synthetic resins (Fractogel, Toyopearl,
polyacrylamide) for gel filtration of these proteins.
Peeter Toomik

Sham Nair wrote:


> Hi Netters, I am trying to purify a calcium-dependent lectin
> from the serum of an invertebrate.  After initial affinity
> chromatography (using N-acetyl-D-galactosamine coupled to
> agarose and eluting with EDTA), I tried to separate the
> mixture of proteins in the eluate by size exclusion
> chromatography.  The problem was that I lost the lectin during
> this latter process.  I presumed that the lectin was binding
> to the chromatography media (Sepharose) and tried to carry out
> the separation with EDTA in my lectin suspension, but this did
> not help.  Any suggestions?  Thanks in advance.
>
> Regards,
> Sham
>
>