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Separation of basic drug enantiomers by capillary electrophoresis using chicken α 1 ‐acid glycoprotein: Insight into chiral recognition mechanism
Author(s) -
Matsunaga Hisami,
Sadakane Yutaka,
Haginaka Jun
Publication year - 2003
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.200305483
Subject(s) - recombinant dna , orosomucoid , chemistry , alpha (finance) , enantiomer , affinity electrophoresis , chromatography , capillary electrophoresis , microbiology and biotechnology , biochemistry , glycoprotein , affinity chromatography , enzyme , stereochemistry , biology , gene , medicine , construct validity , nursing , patient satisfaction
Abstract Recombinant chicken α 1 ‐acid glycoprotein (α 1 ‐AGP) was prepared by the Escherichia coli expression system and completely deglycosylated α 1 ‐AGP (cd‐α 1 ‐AGP) was obtained by treatments of native α 1 ‐AGP with a mixture of endoglycosidase and N ‐glycosidase. The average molecular masses of chicken α 1 ‐AGP, cd‐α 1 ‐AGP and recombinant α 1 ‐AGP were estimated to be about 29 200, 21 700 and 20 700, respectively, by matrix‐assisted laser desorption‐time of flight‐mass spectrometry. We compared the chiral recognition ability of chicken α 1 ‐AGP, cd‐α 1 ‐AGP and recombinant α 1 ‐AGP using them as chiral selectors in capillary electrophoresis. The chicken α 1 ‐AGP showed higher resolution for eperisone, pindolol and tolperisone than cd‐α 1 ‐AGP or recombinant α 1 ‐AGP. Recombinant α 1 ‐AGP still showed chiral recognition for three basic drugs tested. By addition of propranolol as a competitor in the separation solution in CE, no enantioseparations of three basic drugs were observed with chicken α 1 ‐AGP, cd‐α 1 ‐AGP or recombinant α 1 ‐AGP. These results reveal that the protein domain of the chicken α 1 ‐AGP is responsible for the chiral recognition ability, and that the chiral recognition site(s) for basic drugs exists on the protein domain.