[RASMB] High sucrose concentrations in sed velocity experiments

Parker, Matthew Matthew.Parker at immunogen.com
Mon Nov 14 07:24:32 PST 2005


            Hi. We recently ran sedimentation velocity on some samples that
contained 10% sucrose in the buffer. We obtained results for the c(s)
distribution that didn't agree well with what we saw by SEC, and the s-values
were also shifted to significantly lower values than what we would expect
(and that we have seen in other formulations) for an IgG. It was suggested to
us that our sucrose concentration was high enough to create a sucrose
gradient in the cell during the course of the run, and that this could cause
the relative peak areas in the c(s) plot to be inaccurate.

 

            Does anybody have any comments about this, or suggestions about
how high in sucrose content you can go before this starts to become a
problem?

 

            Thanks,

 

            Matt

 

Matthew Parker, Ph.D.

Analytical and Pharmaceutical Sciences

Immunogen, Inc.

Cambridge, MA

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