Matt, there should be also a material called PBE94 for pH range 9 to 4
(and another one called Pharmalyte for high pH) to pack your own column.
It worked pretty well for our protein and is not in the high price
region of the MonoP column.
Best,
Guenter
Matthew Chu wrote:
> Thanks Guenter and Andreas,
>
> Yea, I have taken a look for the Mono P before, I thought the material
> they used in Mono P is basically the same as in Mono Q and I found the
> book "Protein Purification Protocols: Second Edition" by Paul Cutler
> mentioned that phosphate buffer can also generate a continuous
> gradient....that's why I reckon we can also perform pH gradient in
> Mono Q. But I am worrying if it is a good idea to go for it, as both
> mono Q or mono P are quite expensive.
>
> Matt
>
> 2008/6/24 Andreas Förster <[log in to unmask]
> <mailto:[log in to unmask]>>:
>
> Hey Matt,
>
> it seems to me that what you're asking for is chromatofocusing.
> See the official GE documentation:
> http://www6.gelifesciences.com/aptrix/upp00919.nsf/Content/WD:Chromatofocusin(260949098-R350)
> <http://www6.gelifesciences.com/aptrix/upp00919.nsf/Content/WD:Chromatofocusin%28260949098-R350%29>
>
> The proprietary buffers are a bit expensive, but as you found out,
> they're a bit complicated to make. I don't know if you'd ruin
> your Mono Q with a pH gradient. If in doubt, buy one of the
> dedicated Mono P column.
>
> Hope that helps.
>
>
> Andreas
>
>
> Matthew Chu wrote:
>
> Dear All,
>
> Sorry for off-topic question. Does anyone have any experience
> in purifying protein using pH gradient in Mono Q column?
>
> I have been googling for a whole day, only one paper was found
> to mention performing pH gradient in Mono Q, but in a mixture
> of amine buffering species, which is a bit too complicated (J.
> Chromatogr. A 1164 (2007) 181 - 188. Can Tris-Cl/Tris-base or
> phosphate buffer give a linear pH gradient from pH 8.0 to 4.0?
> Is it usual to perform pH gradient in Mono Q as I don't want
> to ruin my Mono Q column...
>
> Any suggestions are welcome. Thanks in advance!
>
> Kind regards,
> Matt
>
> ----------------------------------------------------------------------------
> Matthew LH Chu
> PhD Student
> School of Pharmacy and Pharmaceutical Sciences
> University of Manchester
> ----------------------------------------------------------------------------
>
>
>
>
> --
> ----------------------------------------------------------------------------
> Matthew LH Chu
> PhD Student
> School of Pharmacy and Pharmaceutical Sciences
> University of Manchester
> ----------------------------------------------------------------------------
--
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