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Effect of pore size on performance of monolithic tube chromatography of large biomolecules
Author(s) -
Podgornik Ales,
Hamachi Masataka,
Isakari Yu,
Yoshimoto Noriko,
Yamamoto Shuichi
Publication year - 2017
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201700258
Subject(s) - monolith , pressure drop , chromatography , chemistry , volumetric flow rate , analytical chemistry (journal) , drop (telecommunication) , ion chromatography , biomolecule , packed bed , size exclusion chromatography , materials science , thermodynamics , organic chemistry , telecommunications , biochemistry , physics , computer science , enzyme , catalysis
Effect of pore size on the performance of ion‐exchange monolith tube chromatography of large biomolecules was investigated. Radial flow 1 mL polymer based monolith tubes of different pore sizes (1.5, 2, and 6 μm) were tested with model samples such as 20 mer poly T‐DNA, basic proteins, and acidic proteins (molecular weight 14 000–670 000). Pressure drop, pH transient, the number of binding site, dynamic binding capacity, and peak width were examined. Pressure drop–flow rate curves and dynamic binding capacity values were well correlated with the nominal pore size. While duration of the pH transient curves depends on the pore size, it was found that pH duration normalized on estimated surface area was constant, indicating that the ligand density is the same. This was also confirmed by the constant number of binding site values being independent of pore size. The peak width values were similar to those for axial flow monolith chromatography. These results showed that it is easy to scale up axial flow monolith chromatography to radial flow monolith tube chromatography by choosing the right pore size in terms of the pressure drop and capacity.