Transfection reagents

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Tue Nov 30 02:50:04 EST 1999



Benny Lo wrote:

> Dear All
>
> I 've been trying to transfect some COS-7 cell lines with an expression
> vector containing the protein of interest. I've tried using electroporation
> but has had no luck. Besides electroporation, I 've also tried using
> SuperFect from Qiagen. Does anyone have any more experience with
> transfection reagents as to which is the best, bearing in mind that my goal
> is to produce proteins from COS-7 cells (and subsequently CHO cells) for
> purification so the possibility of scaling up is preferable.
>
> Any advice will be appreciated. Many thanks.
>
> Rgds
> Benny.

Hi Benny,
There are quite a number of transfection methods available that introduce your
expression vector into cells. COS-7 cells are easily transfected with most of
these methods, but other cells are much more difficult. If your goal is to
scale up the cell culture for purification of a recombinant protein, it is
important that you use a method that is affordable in larger scales. All the
"modern" materials, including Superfect, work great but are very expensive.
There are two options that may be worth a try. You could either use
Ca-phosphate transfection for transient transfections (even if you won´t get
more than 10-20% transfected cells) or make a stable transfection to get a
producing cell line. Making cell lines is easy but takes quite some time (2
months), so you have to decide if it is worth the time and effort in your
particular case.

Hope this helps,
Frank





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