Purifying Antibodies using affinity support

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Tue Mar 9 10:56:19 EST 1999


In article <36E524C9.5971A872 at arches.uga.edu>, Keith Pitts <pitts at arches.uga.edu> wrote:
>Hey-
>
>Our lab needs to clean up a weak antibody that has a relatively low
>titer to our protein.  It has been suggested that we use a method
>whereby we covalently attach the purified protein to a resin and
>affinity purify the antibodies to see if we can improve the
>specificity.  So far, I have found a Bio-Rad support gel which will bind
>primary amines (which may be fine for our purposes), but does anyone
>know much about this process?  Also does anyone know of any other _good_
>supports to use for this endeavor?
>

'Good' in this case is always relative. Some work best in some cases,
some in others. Bio-Rad Affi Gel (forgot the number) is NHS activated
support with relatively low capacity. It is good. The most common alternative
is to use bromcyan avtivated Sepharose. There are others, less popular
chemistries. Pierce sells what appears to be very attractive new approach
(I think it's called Emphase?). 

I personally always use periodate activated CL sepharose - a method I 
learned over 10 years ago when I was student. Cheap, reliable and 
extremely efficient. I am not sure if it ever was published - I tried
to find papers from people who developed it and taught me, but 
did not find any. I can post it if necessary.

        - Dima




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