We are a small company that manufactures multi-angle laser light
scattering detectors (MALLS) used in combination with chromatographically
separated samples to determine their absolute molecular weights and sizes
and the distributions of these quantities. During the past few years, a
great number of our colleagues seem to be increasingly interested in
proteins and their characterizations. (E. g. aggregation phenomena, self
assembly, stability and just plain molecular weights of various
preparation.) We get a lot of complaints, not so much about the
instruments, but about the chromatography itself and what the separation
process is doing to the samples. We're not experts in protein chemistry,
but obviously feel we have a lot to learn both to help our customers and
to interact better with the protein chemistry community. Right now we
have the following four questions and would be most grateful for any
comments or suggestions a protein or biochemist might have:
Please be assured that this is NOT a commercial pitch, but it seems to
touch on some pretty important measurements now going on in the protein
community.
1) What do you think of the chromatographic separation process as
a tool for characterizing protein samples (assuming that an absolute
determination may result)?
2) If you use such techniques, what type do you use? (E. g.
reverse phase, size exclusion, ion exchange, capillary electrophoresis)
3) Do you think the buffer and/or the mobile phase can (will)
affect the composition of your sample? (E. g. might you detect
aggregates that were not present in your sample? Could some of the sample
stick to the columns during the separation process itself?)
4) If a column structure could be developed that would permit
the use of any type of mobile phase without affecting the separation
process itself (i. e. no sticking and no false aggregation), would that
make such chromatographic techniques more attractive? (We do NOT
manufacture columns!)
Phil Wyatt
wyatt at wyatt.com
