Re: [ccp4bb] ultrafiltration/buffer exchange device for insect cell cultures

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

Xtal community:

- CCP4BB

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: refinment in lower space group vs. higher
From: Anastassis Perrakis a {- dot -} perrakis {- at -} NKI {- dot -} NL
Date: 2009-01-05

Next message:
Subject: Re: problem about BALBES
From: Xiang Liu pku06a15 {- at -} YAHOO {- dot -} COM {- dot -} CN
Date: 2009-01-05

Subject: Re: ultrafiltration/buffer exchange device for insect cell cultures
From: "R {- dot -} M {- dot -} Garavito" garavito {- at -} MSU {- dot -} EDU
Date: 2009-01-05

Look into the 0.5 sq. m Millipore Pellicon ultrafiltration/
microfiltration devices (http://www.millipore.com/catalogue/module/
c558). They are moderately expensive, but the membranes are robust,
reusable and allow fast concentration steps. We have regularly used
them to concentrate cells (microfiltration mode) and/or media
(ultrafiltration mode) from 16 L insect cell cultures to volumes of
500 mL in ~1 hr. Given the speed, you could use it for buffer exchange.

Michael

****************************************************************
R. Michael Garavito, Ph.D.
Professor of Biochemistry & Molecular Biology
513 Biochemistry Bldg.
Michigan State University
East Lansing, MI 48824-1319
Office: (517) 355-9724 Lab: (517) 353-9125
FAX: (517) 353-9334 Email: garavito@msu.edu
****************************************************************

On Jan 4, 2009, at 8:34 PM, 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
>

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: refinment in lower space group vs. higher
From: Anastassis Perrakis a {- dot -} perrakis {- at -} NKI {- dot -} NL
Date: 2009-01-05

Next message:
Subject: Re: problem about BALBES
From: Xiang Liu pku06a15 {- at -} YAHOO {- dot -} COM {- dot -} CN
Date: 2009-01-05