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CCP4BB  January 2009

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Subject:

Re: ultrafiltration/buffer exchange device for insect cell cultures

From:

Dean Madden <[log in to unmask]>

Reply-To:

Dean Madden <[log in to unmask]>

Date:

Wed, 7 Jan 2009 18:16:23 -0500

Content-Type:

text/plain

Parts/Attachments:

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text/plain (48 lines)

As an alternative to the Pellicon system, Millipore also makes PrepScale 
TFF cartridges (http://www.millipore.com/techpublications/tech1/ef063). 
Unlike the Pellicon, they are sealed units, but like the Pellicon 
membranes, they are true tangential-flow devices and can be reused many 
times (>20preps or ~1year in our hands). The time required to 
concentrate insect-cell culture medium from 10-20L to <1L, is also on 
the order of hours, depending on the MWCO and the size of the cartridge. 
With <10L, the 2.5ft^2 30000MWCO cartridge should take 1-2 hours. For 
 >10L at 30000MWCO, use the 6ft^2 cartridge.

When we compared prices a number of years ago, the upfront cost was 
lower with the TFF. If you're going to be a long-term heavy user, it may 
be worth it to invest in the Pellicon. Prices may have changed...

Another point: if you use a peristaltic pump like the Masterflex, keep a 
close eye on the tubing in the head. The back pressure can be 
significant at the required flow rates and can cause tubing failure. We 
replace the section within the pump head after each use.

Dean


Michael Hothorn wrote:
> Dear all,
> 
> sorry for the non-CCP4 question. I am expressing a protein in insect 
> cells that is targeted for secretion into the medium. After expression, 
> I want to concentrate the medium containing my protein of interest (from 
> 10 l to 0.3 L) and at the same time change the buffer to something that 
> is compatible with my first ion exchange chromatography step. I would be 
> interested to know what kind of ultrafiltration devices can be used for 
> this purpose, and whether some of you could suggest a quick, reliable 
> and efficient system that we could purchase for our lab.
> 
> Thanks!
> Michael
> 

-- 
Dean R. Madden, Ph.D.
Department of Biochemistry
Dartmouth Medical School
7200 Vail Building, Rm 408A
Hanover, NH 03755-3844 USA

tel: +1 (603) 650-1164
fax: +1 (603) 650-1128
e-mail: [log in to unmask]

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