Direct resolution of enantiomers in high-performance immunoaffinity chromatography under isocratic conditions.
This paper describes the application of stereoselective antibodies as tailor-made chiral selectors for the separation of enantiomers in HPLC under isocratic conditions. Stereoselective monoclonal antibodies to D- and L-alpha-amino acids, raised against protein conjugates of p-amino-D- and L-phenylalanine, were immobilized on a synthetic high-flow-through support material and used for rapid enantiomer separation of a number of amino acids at flow rates between 0.1 and 10 mL/min. Since separations could be performed in a mild buffer, column lifetime considerably exceeded that of classical immunoaffinity systems. Using an anti-D-amino acid antibody as chiral selector, the L-enantiomers eluted with the void volume, while the D-enantiomers eluted second. Inverted elution orders were obtained on chiral stationary phases prepared from an anti-L-amino acid antibody. These results demonstrate, for the first time, that antibody-based chiral stationary phases are useful for routine enantiomer separation under true high-performance chromatographic conditions.
Hofstetter
Oliver
O
Department of Chemistry and Biochemistry, Northern Illinois University, DeKalb 60115, USA. ohofst@niu.edu
Lindstrom
Heather
H
Hofstetter
Heike
H
D000596:Amino Acids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D051379:Mice; D011506:Proteins / Q000737:chemistry / Q000276:immunology; D013237:Stereoisomerism
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D011506:Proteins