关键词: chromatography;protein;review

Abstract: The first dedicated protein chromatography media were introduced during the 1950s and
1960s. There was an early awareness of the possibility of using these for production
applications within the biopharmaceutical industry. However, the crucial limitation was the
fact that those media that were most compatible with proteins lent themselves less
favourably to scaling-up. The problems were primarily physical. Thus the fibrous cellulose
media showed bed cracking tendencies and the bead shaped polyacrylamide, dextran, and
agarose gel media, then available,were too soft to stand the hydrodynamic forces acting in
large columns, leading to bed compaction and increased pressure drop. At the time, the best
solution to the latter problem, after a number of intermediary solutions were tried, was
the introduction of the stacked column concept in which several short column segments were
connected by small bore tubing, thus reducing the force acting on the particles in each bed
compartment. However, the ultimate remedy, the introduction of chromatographic matrices
that combine the desired features of adequate rigidity,macroporosity, biocompatibility,
chemical stability (for CIP and SIP) and derivatizability, did not occur until the middle
of the 1980s when adequately cross-linked agarose gel media such as Sepharose(R) Fast Flow
were made available. The paper also recognizes the many attempts made during the past 50
years to develop continous chromatography columns. Most of the designs are based on an
annular bed or on an array of annularly arranged parallel columns continuously fed with
samples in a cyclic manner. The introduction of media and columns for expanded bed
adsorption followed a demand for fewer purification steps and shorter process times. In
recent years, columns have been introduced that allow packing and repacking without needing
to open the column. The review provides an historical account of the developments that have
led to the present state-of-the-art both regarding large diameter columns and gel media
intended for industrial applications of protein chromatography and also discusses the
current trends that point to possible future applications.

Key words: chromatography, protein, review