Conductivity and pH Monitoring Optimizes Protein Separations
Ion exchange column chromatography has been a key method for
the separation and purification of proteins for over thirty
years. The need for faster separation of large, labile
biomolecules has led to the development of ion exchange HPLC
columns and techniques. The separation must not only be rapid,
but must give high recovery of the component of interest and
must not appreciably affect the biological activity of the
Proteins which are now being separated and purified by ion
exchange include monoclonal antibodies and interferons, viral
proteins, and various intracellular proteins.
Both the classical column and the new HPLC methods require a
simple, yet reliable and reproducible, technique for tracking
the protein of interest as it elutes from the column.
Experience has taught that this tracking can be accomplished
by monitoring eluent conductivity during the separations,
since a protein will usually elute from the column at the same
conductivity value run after run.
Flow through conductivity monitors required to fill this need
have in the past either been very expensive or have lacked the
desired dead volume or conductivity range.
Lazar Research Laboratories, Inc. has introduced an economical
flow through conductivity monitor (Model ICM-180) which can
help optimize the separation and recovery of proteins using
both classical column and HPLC systems.
The conductivity cell attaches to the eluent end of the column
and digitally displays conductivity during a run. It has a
dead volume of less than 20 microliters, insuring that no
remixing of the separated components will occur as well as
being totally biocompatible.
It also has a broad conductivity range to accommodate most
salt gradients and attaches directly to any strip chart
recorder for a record of the conductivity profile. The
conductivity monitor is also useful for the quick regeneration
of ion exchange columns. Lazar has also recently introduced a
flow through pH monitor (Model FTPH-1A) for chromafocusing
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