Handbook of Size Exclusion Chromatography and Related ...

into accepted laboratory techniques through the availability of commercial
supplies of agarose- and polystyrene-based packing materials.
During the initial stages of development, the particle size of SEC packings
did not decrease as rapidly as that of silica-based packings employed in highperformance
liquid chromatography (HPLC) techniques. According to theory,the
performance of HPLC columns improves in direct proportion to adecrease in
particle size (3). This prediction was proven correct during the latter part of the
1960s. It was not until the late 1970s, however, that this concept led to the use of
smallsilica-basedparticlesforsizeexclusionchromatographysupports.The5-mm
silicagelparticleswerefirstshowntobeanefficientsubstitutefortraditionalresinbasedparticlesinGPC(4).Later,thepotentialofsilicaandporousglassforusein
GFC was demonstrated, following their chemical bonding with hydrophilic
ligands to prevent adsorption of proteins and nucleic acids (5).
Since their introduction in 1978, high-performance silica-based SEC
packingshavemadeagreatimpactintheanalysisandpurificationofbiopolymers.
Columnsfilledwith10-mmsphericalparticlesandnominalporesizesof125,250,
and 500 A ˚ (10 A ˚ ¼1nm) became the state of the art for protein separations
duringthe1980s(6).Furtherimprovementsinspeedandresolutionwereobtained
byreducingthesizeoftheparticlesfrom10to5mm(7).Columnsfilledwiththese
high-performance particles are now manufactured and distributed by several
companies. Although this chapter discusses several aspects of the use of silicabasedpackingsfor
biopolymer analysis,consultChapters15and16fordetailson
the application of SEC for the separation of proteins and nucleic acids,
respectively.
For the analysis of organic-soluble and water-soluble synthetic polymers,
silica-based packing materials have not become as widely used as was originally
envisioned (8). Major improvements in the properties of polymer-based supports
have contributed to their increased use in GPC. Columns packed with polystyrene
divinylbenzene particles are now as efficient as those filled with silica particles
of the same size. Because polymer-based packings can be synthesized with very
small (,60 A ˚ ) and very large (.4000 A ˚ ) pores, they provide better selectivity
than silica columns for the separation of monomers, as well as for very high
molecular weight (5–20 million dalton) polymers.
The use of (modified) silica gels for size exclusion chromatography has been
the topic of many recent reviews and books. The 1979 book from Yau et al.,
enriched by the authors’ contribution to the development of high-performance
silica-based SEC packings, is still an often-used reference for new and experienced
workers alike (8). The application of silica-based packing materials for biopolymer
separations is discussed in detail in Refs 9–14. References 15–17 focus mainly on
gels (organic nonrigid packing materials), which are exclusively discussed in
Refs 18 and 19. Refer to the comprehensive review from Barth and Boyes (20) for
recent references for the analysis of organic- and water-soluble industrial
© 2004 by Marcel Dekker, Inc.