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Single step synthesis of carbohydrate monolithic capillary columns for affinity chromatography of lectins
Author(s) -
Tetala Kishore K. R.,
Chen Bo,
Visser Gerben M.,
van Beek Teris A.
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700356
Subject(s) - chemistry , chromatography , galactose , carbohydrate , monomer , elution , mannose , methacrylate , affinity chromatography , lectin , biochemistry , organic chemistry , polymer , enzyme
Carbohydrate monolithic beds were synthesized in a single step in capillary columns to study affinity chromatography of lectins. In this method, carbohydrates (β‐galactose, β‐glucose, and α‐mannose) with an easy to synthesize alkene terminated tetraethylene glycol spacer were used as functional monomers along the monomer 2‐hydroxyethyl methacrylate (HEMA). As crosslinkers (+)‐ N , N ′‐diallyltartardiamide (DATD) and piperazine diacrylamide (PDA, 1,4‐bisacryloyl‐piperazine) were used. SEM showed the successful formation of monolithic beds in the capillary columns. The permeability of the columns was high. The specific interaction of the lectins Con A, Lens culinaris (LCA) and Arachis hypogaea (PNA) with the carbohydrate stationary phase was studied by frontal affinity chromatography (FAC). Con A and LCA were successfully eluted from the column using 0.1 M methyl‐α‐mannopyranoside and PNA with 0.1 M β‐galactose. Dissociation constants ( K d ) for carbohydrate–lectin interactions were determined and compared with literature.