Biomedical Engineering Reference
In-Depth Information
Figure 6.18
Preparative HPLC column (15 cm in diameter) used in processing of proteins required for thera-
peutic or diagnostic purposes. Column manufactured by Prochrom, Nancy, France (photograph courtesy of
Affi nity Chromatography Ltd)
in downstream processing of low-volume, extremely high-value proteins, mostly intended for ther-
apeutic use, as opposed to proteins used for industrial preparations.
An alternative chromatographic system to HPLC is also available. Termed fast protein liquid chro-
matography (FPLC), this technique employs operating pressures signifi cantly lower than those used in
conventional HPLC systems. Lower pressures allow the use of matrix beads based on polymers such
as agarose. FPLC columns are constructed of glass or inert plastic materials. Conventional HPLC col-
umns are manufactured from high-grade stainless steel. In many cases, FPLC systems are economi-
cally more attractive than HPLC. Despite their operation at lower pressures, they still combine high
resolution with enhanced speed of operation when compared with traditional low-pressure systems.
6.8 Purifi cation of recombinant proteins
Proteins produced by recombinant DNA technology are usually purifi ed by means identical to
those available for purifi cation of traditional non-recombinant proteins extracted directly from
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